Genetic Counseling
Chapter
13
Outline
	13.1	Counseling for Chromosomal Disorders
	•	A karyotype is a visual display of an individual’s chromosomes arranged by pairs.196
	•	Amniocentesis and chorionic villi sampling provide fetal cells for karyotyping.197
	•	Chromosomal mutations can be due to a change in either number or structure.198
	13.2	Counseling for Genetic Disorders: The Present
	•	Constructing a pedigree shows the pattern of gene inheritance and the chances a genetic disorder will be passed on.200
	•	Several genetic disorders are due to changes in single genes carried on either the autosomes or the X chromosome.200–201
	•	It is possible to test the individual, the fetus, the embryo, or the egg for a mutated gene.205–7
	13.3	Counseling for Genetic Disorders: The Future
	•	The Human Genome Project has revealed the sequence of all the base pairs in the human genome.208
	•	DNA chips (DNA microarrays) contain the entire human genome and can be used to create a genetic profile of an individual.209
	•	Genetic profiles will lead to better preventive care, drug therapies, and gene therapy.209
	13.4	Gene Therapy
	•	Ex vivo techniques involve genetic engineering of a patient’s cells outside the body.210
	•	In vivo techniques involve attempting to alter cells without removing them.211
Today it is possible to screen embryos for genetic abnormalities before they are implanted in the uterus. This process is called pre-implantation genetic diagnosis (PGD). Couples who are at high risk of having a child with a genetic disorder can use PGD to increase their chances of having a normal child. The couple donates a number of egg cells and sperm cells, and the embryos are created in a lab by a process called in vitro fertilization. Once the embryos have enough cells, one cell from each embryo can be removed and tested for genetic mutations. If an embryo is found to have a mutation for a genetic disorder of concern, it can be discarded. Embryos that appear free of genetic flaws can be implanted into the mother. If all goes well, a healthy baby will be born.
Some people have ethical concerns about using PGD to select genetically healthy embryos. Even more controversial, however, is the use of this technology purely to choose the sex of an embryo. Some couples so desperately want a boy or a girl that they use PGD to guarantee the gender of their child, and some physicians justify this use of the technology for the achievement of “gender-balance” within a family. This use of PGD is legal in the United States, but has been banned in some countries.  
In this chapter, you will learn about some of the techniques currently being used to counsel couples about their chances of passing on a genetic disorder, as well as technologies that may be used to test and treat parents and children for genetic disorders in the future. 
13.1		Counseling for Chromosomal Disorders
Potential parents are becoming aware that many illnesses are caused by abnormal chromosomal inheritance or by gene mutations. Therefore, more couples are seeking genetic counseling, which is available at many major hospitals as a means to determine the risk of inherited disorders in a family. For example, a couple might be prompted to seek counseling after several miscarriages, when several relatives have a particular medical condition, or if they already have a child with a genetic defect. The counselor helps the couple understand the mode of inheritance, the medical consequences of a particular genetic disorder, and the decisions they might wish to make (Fig. 13.1). 
Various human disorders result from abnormal chromosome number or structure. When a pregnant woman is concerned that her unborn child might have a chromosomal defect, the counselor may recommend karyotyping the fetus’s chromosomes. 
Karyotyping
A karyotype is a visual display of the chromosomes arranged by size, shape, and banding pattern. Any cell in the body except red blood cells, which lack a nucleus, can be a source of chromosomes for karyotyping. In adults, it is easiest to use white blood cells separated from a blood sample for this purpose. In fetuses, whose chromosomes are often examined to detect a syndrome, cells can be obtained by either amniocentesis or chorionic villi sampling.
Amniocentesis is a procedure for obtaining a sample of amniotic fluid from the uterus of a pregnant woman. Blood tests and the age of the mother are considered when determining whether the procedure should be done. The risk of spontaneous abortion increases by about 0.3% due to amniocentesis, and doctors use the procedure only if it is medically warranted.
Amniocentesis is not usually performed until about the fourteenth to the seventeenth week of pregnancy. A long needle is passed through the abdominal and uterine walls to withdraw a small amount of fluid, which also contains fetal cells (Fig. 13.2a). Tests are done on the amniotic fluid. Karyotyping the chromosomes may be delayed as long as four weeks so that the cells can be cultured to increase their number.
Chorionic villi sampling (CVS) is a procedure for obtaining chorionic cells in the region where the placenta will develop. This procedure can be done as early as the fifth week of pregnancy. A long, thin suction tube is inserted through the vagina into the uterus (Fig. 13.2b). Ultrasound, which gives a picture of the uterine contents, is used to place the tube between the uterine lining and the chorionic villi. Then, a sampling of chorionic cells is obtained by suction. The cells do not have to be cultured, and karyotyping can be done immediately. But testing amniotic fluid is not possible because no amniotic fluid is collected. Also, CVS carries a greater risk of spontaneous abortion than amniocentesis—0.8% compared to 0.3%. The advantage of CVS is getting the results of karyotyping at an earlier date.
After a cell sample has been obtained, the cells are stimulated to divide in a culture medium. A chemical is used to stop mitosis during metaphase when chromosomes are the most highly compacted and condensed. The cells are then killed, spread on a microscope slide, and dried. Stains are applied to the slides, and the cells are photographed. Staining causes the chromosome to have dark and light crossbands of varying widths, and these can be used, in addition to size and shape, to help pair up the chromosomes. Today, technicians use a computer to arrange the chromosomes in pairs. As described in Chapter 9, the karyotype of a person who has Down syndrome usually has three number 21 chromosomes instead of two (Fig. 13.2c). 
Chromosomal Mutations
A mutation is a permanent genetic change. A change in chromosome number or structure that can be detected microscopically is a chromosomal mutation. A karyotype reveals changes in chromosome number, and a skilled technician is able to detect differences in chromosome structure based on a change in the normal banding patterns of the chromosomes (see Fig. 13.2c). 
In Chapter 9, you learned about syndromes that result from changes in chromosome number, including Down syndrome, Turner syndrome, and Klinefelter syndrome.
Syndromes that result from changes in chromosome structure are due to the breakage of chromosomes and their failure to reunite properly. Various environmental agents—radiation, certain organic chemicals, or even viruses—can cause chromosomes to break apart. Ordinarily, when breaks occur in chromosomes, the segments reunite to give the same sequence of genes. But their failure to do so results in one of several types of mutations: deletion, duplication, translocation, or inversion. Chromo-somal mutations can occur during meiosis, and if the offspring inherits the abnormal chromosome, a syndrome may develop.
Deletions and Duplications
A deletion occurs when a single break causes a chromosome to lose an end piece, or when two simultaneous breaks lead to the loss of an internal chromosome segment. An individual who inherits a normal chromosome from one parent and a chromosome with a deletion from the other parent no longer has a pair of alleles for each trait, and a syndrome can result.
Williams syndrome occurs when chromosome 7 loses a tiny end piece (Fig. 13.3). Children who have this syndrome look like pixies, with turned-up noses, wide mouths, a small chin, and large ears. Although their academic skills are poor, they exhibit excellent verbal and musical abilities. The gene that governs the production of the protein elastin is missing, and this affects the health of the cardiovascular system and causes their skin to age prematurely. Such individuals are very friendly but need an ordered life, perhaps because of the loss of a gene for a protein that is normally active in the brain.
Cri du chat (cat’s cry) syndrome occurs when chromosome 5 is missing an end piece. The affected individual has a small head, is mentally retarded, and has facial abnormalities. Abnormal development of the glottis and larynx results in the most characteristic symptom—the infant’s cry resembles that of a cat.
In a duplication, a chromosome segment is repeated so that the individual has more than two alleles for certain traits. An inverted duplication is known to occur in chromosome 15. (Inversion means that a segment joins in the direction opposite from normal.) Children with this syndrome, called inv dup 15 syndrome, have poor muscle tone, mental retardation, seizures, a curved spine, and autistic characteristics, including poor speech, hand flapping, and lack of eye -contact (Fig. 13.4).
Translocation
A translocation is the exchange of chromosome segments between two nonhomologous chromosomes. A person who has both of the involved chromosomes has the normal amount of genetic material and is healthy, unless the chromosome exchange breaks an allele into two pieces. The person who inherits only one of the translocated chromosomes will no doubt have only one copy of certain alleles and three copies of other alleles. A genetic counselor begins to suspect a translocation has occurred when spontaneous abortions are commonplace and family members suffer from various syndromes. A special microscopic technique allows a technician to determine that a translocation has occurred.
In 5% of Down syndrome cases, a translocation that occurred in a previous generation between chromosomes 21 and 14 is the cause. As long as the two chromosomes are inherited together, the individual is normal. But in future generations a person may inherit two normal chromosomes 21 and the abnormal chromosome 14 that contains a segment of chromosome 21. In these cases, Down syndrome is not related to parental age but instead tends to run in the family of either the father or the mother.
Figure 13.5 shows a father and son who have a translocation between chromosomes 2 and 20. Although they have the normal amount of genetic material, they have the distinctive face, abnormalities of the eyes and internal organs, and severe itching characteristic of Alagille syndrome. People with this syndrome ordinarily have a deletion on chromosome 20; therefore, it can be deduced that the translocation disrupted an allele on chromosome 20 in the father. The symptoms of Alagille syndrome range from mild to severe, so some people may not be aware they have the syndrome. This father did not realize it until he had a child with the syndrome.
Inversion
An inversion occurs when a segment of a chromosome is turned 180°. You might think this is not a problem because the same genes are present, but the reverse sequence of alleles can lead to altered gene activity.
Crossing-over between an inverted chromosome and the noninverted homologue can lead to recombinant chromosomes that have both duplicated and deleted segments. This happens because alignment between the two homologues is only possible when the inverted chromosome forms a loop (Fig. 13.6).
13.2		Counseling for Genetic Disorders: The Present
Even if no chromosomal abnormality is likely, amniocentesis still might be done because it is now possible to perform biochemical tests on amniotic fluid to detect over 400 different disorders caused by specific genes in a fetus. The genetic counselor determines ahead of time what tests might be warranted. To do this, the counselor needs to know the medical history of the family in order to construct a pedigree.
Family Pedigrees
A pedigree is a chart of a family’s history with regard to a particular genetic trait. In the chart, males are designated by squares and females by circles. Shaded circles and squares are affected individuals; they have a genetic disorder. A line between a square and a circle represents a union. A vertical line going downward leads directly to a single child; if there are more children, they are placed off a horizontal line.
From the counselor’s knowledge of genetic disorders, he or she might already know the pattern of inheritance of a trait—that is, whether it is autosomal dominant, autosomal recessive, or X-linked recessive. The counselor can then determine the chances that any child born to the couple will have the abnormal phenotype.
Pedigrees for Autosomal Disorders
A family pedigree for an autosomal recessive disorder is shown in Figure 13.7. In this pattern, a child can be affected when neither parent is affected. Such heterozygous parents are carriers because, although they are unaffected, they are capable of having a child with the genetic disorder. If the family pedigree suggests that the parents are carriers for an autosomal recessive disorder, the counselor might suggest confirming this by doing the appropriate genetic test. Then, if the parents so desire, it would be possible to do prenatal testing of the fetus for the genetic disorder. 
Figure 13.7 lists other ways that a counselor may recognize an autosomal recessive pattern of inheritance. Notice that in this pedigree, cousins are the parents of three children, two of whom have the disorder. Aside from illustrating that reproduction between cousins is more likely to bring out recessive traits, this pedigree also shows that “chance has no memory”; therefore, each child born to heterozygous parents has a 25% chance of having the disorder. In other words, if a heterozygous couple has four children, each child might have the condition.
Figure 13.8 shows an autosomal dominant pattern of inheritance. In this pattern, a child can be unaffected even when the parents are heterozygous and therefore affected. Figure 13.8 lists other ways to recognize an autosomal dominant pattern of inheritance. This pedigree illustrates that when both parents are unaffected, all their children are unaffected. Why? Because neither parent has a dominant gene that causes the condition to be passed on.
Pedigrees for Sex-Linked Disorders
Figure 13.9 gives a pedigree for an X-linked recessive disorder. Recall that sons inherit an X-linked recessive allele from their mothers because their fathers give them a Y chromosome. More males than females have the disorder because recessive alleles on the X chromosome are always expressed in males—the Y chromosome lacks an allele. Females who have the condition inherited the allele from both their mother and their father, and all the sons of such a female will have the condition. 
If a male has an X-linked recessive condition, his daughters are all carriers even if his partner is normal. Therefore, X-linked recessive conditions often pass from grandfather to grandson. Figure 13.9 lists other ways to recognize an X-linked recessive disorder.
Only a few traits are known to be X-linked dominant. If a condition is X-linked dominant, daughters of affected males have a 100% chance of having the condition. Females can pass an X-linked dominant allele to both sons and daughters. If a female is hetero-zygous and her partner is normal, each child has a 50% chance of escaping an X-linked dominant disorder, depending on which of the mother’s X chromosomes is inherited. 
A few genetic disorders involve genes carried on the Y chromosome. One gene, called the SRY-determining region, is involved in determining gender during development. Others code for membrane proteins, including an enzyme that regulates the movement of ADP into and ATP out of mitochondria. A counselor would recognize a Y-linked pattern of inheritance because Y-linked disorders are present only in males and are passed directly from father to all sons but not to daughters. (Can you explain why?)
Genetic Disorders of Interest
Medical genetics has traditionally focused on disorders caused by single gene mutations, and we will discuss a few of the better known examples. 
Autosomal Disorders
Autosomal disorders are caused by mutated alleles on the autosomal chromosomes (all the chromosomes except the sex chromosomes). Some of these are recessive, and therefore two affected alleles are necessary before an individual has the disorder. Others are dominant, meaning that it takes only one affected allele to cause the disorder. Dominant and recessive inheritance was discussed in Chapter 10.
Tay-Sachs DiseaseTay-Sachs disease is a well-known autosomal recessive disorder that occurs usually among Jewish people of central and eastern European descent. Tay-Sachs disease results from a lack of the enzyme hexosaminidase A (Hex A) and the subsequent storage of its substrate, a glycosphingolipid, in lysosomes. Lysosomes build up in many body cells, but the primary sites of storage are the cells of the brain, which accounts for the onset of symptoms and the progressive deterioration of psychomotor functions (Fig. 13.10).
At first, it is not apparent that a baby has Tay-Sachs disease. However, development begins to slow down between four months and eight months of age, and neurological impairment and psychomotor difficulties then become apparent. The child gradually becomes blind and helpless, develops uncontrollable seizures, and eventually becomes paralyzed and dies.
Cystic FibrosisCystic fibrosis is an autosomal recessive disorder that occurs among all ethnic groups, but it is the most common lethal genetic disorder among Caucasians in the United States. Research has demonstrated that chloride ions (Cl2) fail to pass through a plasma membrane channel protein in the cells of these patients. Ordinarily, after chloride ions have passed through the membrane, sodium ions (Na1) and water follow. It is believed that lack of water then causes abnormally thick mucus in the bronchial tubes and pancreatic ducts, thus interfering with the function of the lungs and pancreas. To ease breathing in affected children, the thick mucus in the lungs must be loosened periodically but still the lungs become infected frequently (Fig. 13.11). Clogged pancreatic ducts prevent digestive enzymes from reaching the small intestine, and to improve digestion, patients take digestive enzymes mixed with applesauce before every meal.
PhenylketonuriaPhenylketonuria (PKU) is an autosomal recessive metabolic disorder that affects nervous system development. Affected individuals lack an enzyme that is needed for the normal metabolism of the amino acid phenylalanine, which therefore appears in the urine and the blood. Newborns are routinely tested in the hospital for elevated levels of phenylalanine in the blood. If elevated levels are detected, newborns will develop normally as long as they are limited to a diet low in phenylalanine until the brain is fully developed, usually around the age of seven. Otherwise, severe mental retardation develops. Some doctors recommend that the diet continue for life, but in any case, a pregnant woman with phenylketonuria must also be on the diet in order to protect her unborn child from harm.
Sickle Cell DiseaseSickle cell disease is an autosomal recessive disorder in which the red blood cells are not biconcave disks like normal red blood cells, but are irregular in shape (Fig. 13.12). In fact, many are sickle-shaped. The defect is caused by an abnormal hemoglobin molecule that differs from normal hemoglobin by one amino acid in the protein globin. The single amino acid change causes hemoglobin molecules to stack up and form insoluble rods, and the red blood cells become sickle-shaped. 
Because sickle-shaped cells can’t pass along narrow capillary passageways as disk-shaped cells can, they clog the vessels and break down. This is why persons with -sickle cell disease suffer from poor circulation, anemia, and low resistance to infection. Internal hemorrhaging leads to further complications, such as jaundice, episodic pain in the abdomen and joints, and damage to internal organs.
 Sickle cell heterozygotes have the sickle cell trait, in which the blood -cells are normal, unless they experience dehydration or mild oxygen deprivation. Still, at present, most experts believe that persons with the sickle cell trait do not need to restrict their physical activity.
Marfan SyndromeMarfan syndrome (see Chapter 10, page 151), an autosomal dominant disorder, is caused by a defect in an elastic connective tissue protein called fibrillin. This protein is normally abundant in the lens of the eye; the bones of limbs; fingers, and ribs; and the wall of the aorta. Thus, the affected person often has a dislocated lens, long limbs and fingers, and a caved-in chest. The aorta wall is weak and can possibly burst without warning. A tissue graft can strengthen the aorta, but affected individuals still should not overexert themselves.
Huntington DiseaseHuntington disease is a dominant neurological disorder that leads to progressive degeneration of neurons in the brain (Fig. 13.13). The disease is caused by a single mutated copy of the gene for a protein called huntingtin. Most patients appear normal until they are of middle age and have already had children, who may then also eventually be stricken. Occasionally, the first sign of the disease in the next generation is seen in teenagers or even younger children. There is no effective treatment, and death comes 10 to 15 years after the onset of symptoms.
Several years ago, researchers found that the gene for Huntington disease was located on chromosome 4. A test was developed for the presence of the gene, but few people want to know if they have inherited the gene because there is no cure. But now we know that the disease stems from a mutation that causes the huntingtin protein to have too many copies of the amino acid glutamine. The normal version of huntingtin has stretches of between 10 and 25 glutamines. If huntingtin has more than 36 glutamines, it changes shape and forms large clumps inside neurons. Even worse, it attracts and causes other proteins to clump with it. One of these proteins, called CBP, helps nerve cells survive. Researchers hope to combat the disease by boosting CBP levels.
Incomplete DominanceIn familial hypercholesterolemia (FH), the liver in homozygotes with two mutated alleles completely lacks low-density lipoprotein (LDL) receptors that take up cholesterol from the bloodstream. Heterozygotes have half the normal number of cholesterol receptors. The number of cholesterol receptors inversely parallels the amount of cholesterol in the plasma. Figure 13.14 compares the amount of plasma cholesterol in the general population with that of heterozygotes and homozygotes for FH. Heart disease occurs in both heterozygotes and homozygotes. Homozygotes die of heart disease as children. Heterozygotes may die when they are young or after they have reached middle age.
X-Linked Recessive Disorders
As discussed in Chapter 10, X-linked recessive disorders are caused by mutated alleles on the X chromosome. A son inherits an X-linked recessive condition from his mother.
Color BlindnessColor blindness is a common X-linked recessive disorder. About 8% of Caucasian men have red-green color blindness. Most of them see brighter greens as tans, olive greens as browns, and reds as reddish-browns. A few cannot tell reds from greens at all; they see only yellows, blues, blacks, whites, and grays.
Duchenne Muscular DystrophyDuchenne muscular dystrophy is an X-linked recessive disorder characterized by wasting away of the muscles. The absence of a protein, now called dystrophin, is the cause of the disorder. Much investigative work determined that dystrophin is involved in the release of calcium from the sarcoplasmic reticulum in muscle fibers. The lack of dystrophin causes calcium to leak into the cell, which promotes the action of an enzyme that dissolves muscle fibers. When the body attempts to repair the tissue, fibrous tissue forms (Fig. 13.15), and this cuts off the blood supply so that more and more cells die.
Symptoms such as waddling gait, toe walking, frequent falls, and difficulty in rising may appear as soon as the child starts to walk. Muscle weakness intensifies until the individual is confined to a wheelchair. Death usually occurs by age 20; therefore, affected males are rarely fathers. The recessive allele remains in the population through passage from carrier mother to carrier daughter.
As therapy, immature muscle cells can be injected into muscles, and for every 100,000 cells injected, dystrophin production occurs in 30–40% of muscle fibers. 
HemophiliaThere are two common types of hemophilia, an X-linked recessive disorder. Hemophilia A is due to the absence or minimal presence of a clotting factor known as factor VIII, and hemophilia B is due to the absence of clotting factor IX. Hemophilia is called the bleeder’s disease because the affected person’s blood either does not clot or clots very slowly. Although hemophiliacs bleed externally after an injury, they also bleed internally, particularly around joints. Hemorrhages can be stopped with transfusions of fresh blood (or plasma) or concentrates of the clotting protein. Also, factors VIII and IX are now available as biotechnology products.
Testing for Genetic Disorders
Following genetic testing, a genetic counselor can explain to prospective parents the chances a child of theirs will have a disorder that runs in their family. If a woman is already pregnant, the parents may want to know whether the unborn child has the disorder. If the woman is not pregnant, the parents may opt for testing of the embryo or egg before she does become pregnant, as described shortly.
Testing depends on the genetic disorder of interest. In some instances, it is appropriate to test for a particular protein, and in others to test for the mutated gene. 
Testing for a Protein 
Some genetic mutations lead to disorders caused by a missing enzyme. For example, in the case of Tay-Sachs disease, it is possible to test for the quantity of the enzyme hex A in a sample of cells and from that, determine whether the individual is likely homozygous normal, a carrier, or has Tay-Sachs disease. If the parents are carriers, each child has a 25% chance of having Tay-Sachs disease. This knowledge may lead prospective parents to opt for testing of the embryo or egg, as discussed on the next page.
In the case of PKU, an enzyme is missing, but the test is performed for the substrate of the enzyme, namely phenylalanine. Paper disks containing the newborn’s blood are placed on a bacterial culture, and if the bacteria grow around them, the newborn has PKU. 
Testing the DNA
Two types of DNA testing are possible: testing for a genetic marker and using a DNA probe.  
Genetic MarkersTesting for a genetic marker relies on a difference in the DNA due to the presence of the abnormal allele. As an example, consider that individuals with sickle cell trait or Huntington disease have an abnormality in a gene’s base sequence. This abnormality in sequence is a genetic marker. As you know, restriction enzymes cleave DNA at particular base sequences. Therefore, the fragments that result from the use of a restriction enzyme will be different for people who are normal than for those who are heterozygous or homozygous for a mutation (Fig. 13.16). 
DNA ProbesA DNA probe is a single-stranded piece of DNA that will bind to complementary DNA. For the purpose of genetic testing, the DNA probe bears a genetic mutation of interest. A new technology that can test for many genetic disorders at a time uses a DNA chip, a very small glass square that contains several rows of DNA probes. Sample DNA is cut into small pieces using restriction enzymes, and the fragments are tagged with a fluorescent dye and converted to single DNA strands before being applied to the chip (Fig. 13.17). Fragments that contain a mutated gene bind to one of the probes, and binding is detected by a laser scanner. Therefore, the results tell whether an individual has particular mutated genes.
Testing the Fetus
If a woman is already pregnant, ultrasound can detect serious fetal abnormalities, and it is also possible to obtain and test the DNA of fetal cells for genetic defects. 
UltrasoundUltrasound images help doctors evaluate fetal anatomy. An ultrasound probe scans the mother’s abdomen, and a transducer transmits high-frequency sound waves that are transformed into a picture on a video screen. This picture shows the fetus inside the uterus (Fig. 13.18). Ultrasound done after 16 weeks can be used to determine the baby’s age and size, and whether there is more than one baby. It’s also possible to tell if a baby has a serious condition, such as spina bifida, which results when the spine fails to close properly during the first month of pregnancy. Surgery to close a newborn’s spine in such a case is generally performed within 24 hours after birth.  
Testing Fetal CellsFetal cells can be tested for various genetic disorders. If the fetus has an incurable disorder, such as Tay-Sachs disease, the parents may wish to consider an abortion. 
For testing purposes, fetal cells may be acquired through amniocentesis or chorionic villi sampling, as described earlier in this chapter. In addition, fetal cells may be collected from the mother’s blood. As early as nine weeks into the pregnancy, a small number of fetal cells can be isolated from the mother’s blood using a cell sorter. While mature red blood cells lack a nucleus, immature red blood cells do have a nucleus, and they also have a shorter life span than mature red blood cells. Therefore, if nucleated fetal red blood cells are collected from the mother’s blood, they are known to be from this pregnancy. 
Only about 1/70,000 blood cells in a mother’s blood are fetal cells, and therefore PCR has to be used to amplify the DNA from the few cells collected. The procedure poses no risk whatsoever to the fetus.
Testing the Embryo and Egg
As discussed in Chapter 29, in vitro fertilization (IVF) is carried out in laboratory glassware. The physician obtains eggs from the prospective mother and sperm from the prospective father, and places them in the same receptacle, where fertilization occurs. Following IVF, now a routine procedure, it is possible to test the embryo. Prior to IVF, it is possible to test the egg for any genetic defect. In any case, only normal embryos are transferred to the uterus for further development. 
Testing the EmbryoIf prospective parents are carriers for one of the genetic disorders discussed earlier, they may want assurance that their offspring will be free of the disorder. Testing the embryo will provide this assurance. 
Following IVF, the zygote (fertilized egg) divides. When the embryo  has six to eight cells, one of these cells can be removed for testing, with no effect on normal development (Fig. 13.19). Only embryos that test negative for the genetic disorders of interest are placed in the uterus to continue developing. 
So far, about 1,000 children worldwide have been born free of alleles for genetic disorders that run in their families following embryo testing. In the future, it’s possible that embryos who test positive for a disorder could be treated by gene therapy, so that they, too, would be allowed to continue to term. 
Testing the EggRecall that meiosis in females results in a single egg and at least two polar bodies. Polar bodies, which later disintegrate, receive very little cytoplasm, but they do receive a haploid number of chromosomes, and thus can be useful in genetic testing. When a woman is heterozygous for a recessive genetic disorder, about half the polar bodies receive the mutated allele, and in these instances the egg receives the normal allele. Therefore, if a polar body tests positive for a mutated allele, the egg received the normal allele (Fig. 13.20). Only normal eggs are then used for IVF. Even if the sperm should happen to carry the mutation, the zygote will, at worst, be heterozygous. But the phenotype will appear normal.
If gene therapy becomes routine in the future, it’s possible that an egg could be given genes that control traits desired by the parents, such as musical or athletic ability, prior to IVF.
13.3		Counseling for Genetic Disorders: The Future
In the previous century, researchers discovered the structure of DNA, how DNA replicates, and how protein synthesis occurs. Genetics in the twenty-first century concerns genomics, the study of genomes (organisms’ genes) to better understand how they direct growth and development and otherwise control the structure and function of the cell. The enormity of this task can be appreciated by knowing that, at the very least, humans have 25,000 genes that code for proteins. An abnormality in any one of these proteins can be the cause of a human illness.
Sequencing the Bases of the Human Genome
We now have a working draft of the base pair sequence in the DNA of all our chromosomes. This feat was accomplished by the Human Genome Project, a 13-year effort that involved both university and private laboratories around the world. How did they do it? First, investigators developed a laboratory procedure that would allow them to decipher a short sequence of base pairs, and then they devised an instrument that would carry out this procedure automatically. As the studies proceeded, DNA sequencers were constantly improved, until today we have instruments that can automatically sequence to 350,000 base pairs of DNA per day (see page 159).  
Genome Comparisons
Researchers are taking all sorts of avenues to link DNA base sequence differences to illnesses. One study compared the human genome to that of chromosome 22 in chimpanzees. Among the many genes that differed in sequence were three of particular interest: a gene for proper speech development, several for hearing, and several for smell. The gene necessary for proper speech development is thought to have played an important role in human evolution. You can suppose that changes in hearing may have facilitated using language for communication between people. Changes in smell genes are a little more problematic. The researchers speculated that the olfaction genes may have affected dietary changes or sexual selection. Or, they may have been involved in traits, other than just smell (Fig. 13.21). It was a surprise to find that many of the other genes they located and studied are known to cause human diseases if abnormal. Perhaps comparing genomes would be a way of finding genes associated with human diseases.
The genomes of many other organisms, such as a common bacterium, a form of yeast, and a species of mouse, are also in the final-draft stage. There are many similarities between the sequences of DNA bases in humans and in other organisms. From this, we can conclude that we share a large number of genes with much simpler organisms, including bacteria! However, with a genome size of 3 billion base pairs, there are also many differences. 
Genetic Profiling 
DNA chips (also called DNA microarrays) can rapidly identify an individual’s complete genotype, including all the various mutations. This genotype is called the person’s genetic profile. To get a genetic profile is easy. The patient provides a few cells, often by simply swabbing the inside of a cheek. The DNA is removed from the cells, amplified by PCR if need be, and cut into fragments, which are tagged by a fluorescent dye. A technician then applies the fragments to a DNA chip, and reads the results. 
With the help of a genetic counselor, individuals can be educated about their genetic profile. It’s possible that a person has or will have a genetic disorder caused by a single pair of alleles. However, polygenic traits are more common, and in these instances, the genetic profile can indicate an increased or decreased risk for a disorder. Risk information can be used to design a program of medical surveillance and to foster a lifestyle aimed at reducing the risk. For example, suppose an individual has mutations common to people with colon cancer. It would be helpful to have an annual colonoscopy so that any abnormal growths can be detected and removed before they become invasive. 
Proteomics and Informatics
The genetic profile may also indicate what drug therapy might be most appropriate for an individual. Because drugs tend to be proteins or small molecules that affect the behavior of proteins, the study of protein function is essential to the discovery of better drugs. One day it may be possible to correlate drug treatment to an individual genetic profile to increase efficiency and decrease side effects. 
The field of proteomics is especially pertinent because it deals with the development of new drugs for the treatment of genetic disorders. Proteomics explores the structure, function, and interaction of cellular proteins. The known sequence of bases in the human genome predicts that at least 25,000 genes are translated into proteins. The translation of all of these genes results in a collection of proteins called the human proteome. Computer modeling of the three-dimensional shape of these proteins is an important part of proteomics. Because the primary structure of these proteins is now known, it should be possible to predict their final shape.
Bioinformatics is the application of computer technologies to the study of the genome (Fig. 13.22). Genomics and proteomics produce raw data, and these fields depend upon computer analysis to find significant patterns in the data. As a result of bioinformatics, scientists hope to find cause-and-effect relationships between various genetic profiles and genetic disorders caused by polygenes.
Also, the current genome sequence contains 82 gene “deserts,” with no known function. Bioinformatics might find that these regions have functions by correlating any sequence changes with resulting phenotypes. New computational tools will most likely be needed in order to accomplish these goals. 
13.4		Gene Therapy
Gene therapy is the insertion of genetic material into human cells for the treatment of a disorder. It includes procedures that give a patient healthy genes to make up for faulty genes, as well as the use of genes to treat various other human illnesses, such as cardiovascular disease and cancer. Gene therapy includes both ex vivo (outside the body) and in vivo (inside the body) methods.
Ex Vivo Gene Therapy
One example of ex vivo gene therapy is the methodology for treating children who have SCID (severe combined immunodeficiency) (Fig. 13.23). These children lack the enzyme ADA (adenosine deaminase), which is involved in the maturation of T and B cells. In order to carry out gene therapy, bone marrow stem cells are removed from the blood and infected with an RNA retrovirus that carries a normal gene for the enzyme. Then the cells are returned to the patient. Bone marrow stem cells are preferred for this procedure because they divide to produce more cells with the same genes. Patients who have -undergone this procedure show significantly improved immune function associated with a sustained rise in the level of ADA enzyme activity in the blood (Fig. 13.24a).
Among the many gene therapy trials, one is for the treatment of familial hypercholesterolemia, described earlier in this chapter as an example of incomplete dominance. High levels of plasma cholesterol make the patient subject to fatal heart attacks at a young age. A small portion of the liver is surgically excised and then infected with a retrovirus containing a normal gene for a cholesterol receptor before the tissue is returned to the patient. Several patients have experienced lowered plasma cholesterol levels following this procedure.
Ex vivo gene therapy is being used in the treatment of cancer. In one procedure, immune system cells are removed from a cancer patient and genetically engineered to display tumor antigens. After these cells are returned to the patient, they stimulate the immune system to kill tumor cells. 
In Vivo Gene Therapy
As mentioned earlier, cystic fibrosis patients lack a gene that codes for a regulator of a transmembrane carrier for the chloride ion. In gene therapy trials, the gene needed to cure cystic fibrosis is sprayed into the nose or delivered to the lower respiratory tract by adeno-viruses or by the use of liposomes (microscopic vesicles that spontaneously form when lipoproteins are put into a solution). Investigators are trying to improve uptake of the genes and are also hypothesizing that a combination of all three vectors might be more successful.
Genes are also being used to treat medical conditions such as poor coronary circulation. It has been known for some time that VEGF (vascular endothelial growth factor) can cause the growth of new blood vessels. The gene that codes for this growth factor can be injected alone, or within a virus, into the heart to stimulate branching of coronary blood vessels. Patients who have received this treatment report that they have less chest pain and can run longer on a treadmill (Fig. 13.24b). 
In vivo gene therapy is increasingly becoming a part of cancer therapy. Genes are being used to make healthy cells more tolerant of chemotherapy, and to make tumors more vulnerable to chemotherapy. The gene p53 brings about apoptosis, and introduction of this gene into cancer cells may be a method of killing them off. 
Figure 13.25 summarizes how gene therapy is being used to treat various illnesses.
THE Chapter in Review
Summary
13.1		Counseling for Chromosomal Disorders
A counselor can detect chromosomal mutations by studying a karyotype of the individual.
Karyotyping
A karyotype is a display of the chromosomes arranged by pairs; the autosomes are numbered from 1 to 22. The sex chromosomes are not numbered.  
Chromosomal Mutations
Nondisjunction during meiosis can result in an abnormal number of autosomes or sex chromosomes in the gametes. Changes in chromosome structure include deletions, duplications, translocations, and inversions: 
In Williams syndrome, one copy of chromosome 7 has a deletion; in cri du chat syndrome, one copy of chromosome 5 has a deletion; and in inv dup 15 syndrome, chromosome 15 has an inverted duplication. Down syndrome can be due to a translocation between chromosomes 14 and 21 in a previous generation.
An inversion can lead to chromosomes that have a deletion and a duplication when the inverted piece loops back to align with the noninverted homologue and crossing-over follows.
13.2		Counseling for Genetic Disorders:
The Present
A counselor can decide the chances of an offspring inheriting a genetic disorder by constructing a pedigree of a disorder that runs in the family.
Family Pedigree  
A family pedigree is a visual representation of the history of a genetic disorder in a family. Constructing pedigrees helps a genetic counselor decide whether a genetic disorder that runs in a family is autosomal recessive (see Fig. 13.7) or dominant (see Fig. 13.8); X-linked (see Fig. 13.9); or some other pattern of inheritance. 
Genetic Disorders of Interest
Autosomal Disorders
•	Tay-Sachs disease (a lysosomal storage disease)
•	Cystic fibrosis (faulty regulator of chloride channel)
•	Phenylketonuria (inability to metabolize phenylalanine)
•	Sickle cell disease (sickle-shaped red blood cells) 
•	Marfan syndrome (defective elastic connective tissue)
•	Huntington disease (abnormal huntingtin protein)
Incomplete Dominance Disorders
•	Familial hypercholesterolemia (liver cells lack cholesterol receptors)
X-Linked Disorders
•	Duchenne muscular dystrophy (absence of dystrophin leads to muscle weakness)  
•	Hemophilia (inability of blood to clot)
Testing for Genetic Disorders
A counselor can order the appropriate test to detect a disorder.
Testing for ProteinsPersons with Tay-Sachs disease lack the enzyme hex A, and those with PKU have much phenylalanine in their blood. 
Testing DNACut DNA with restriction enzymes and then compare the fragment pattern to the normal pattern. See if fragments bind to DNA probes that contain the mutation. 
Testing the FetusAn ultrasound allows for the detection of severe abnormalities, such as spina bifida. Fetal cells can be obtained by amniocentesis or chorionic villi sampling, or by sorting out fetal cells from the mother’s blood. 
Testing the EmbryoFollowing in vitro fertilization (IVF), it is possible to test the embryo. A cell is removed from an 8-celled embryo, and if it is found to be genetically healthy, the embryo is implanted in the uterus, where it develops to term. 
Testing the EggBefore IVF, a polar body can be tested. If the woman is heterozygous, and the polar body has the genetic defect, the egg does not have it. Following fertilization, the embryo is implanted in the uterus. 
13.3		Counseling for Genetic Disorders: 
The Future
In the future, a counselor will be able to study the genetic profile of an individual in order to counsel them about the risk of developing a genetic disorder.
Human Genome ProjectThanks to the Human Genome Project, we now know the sequence of the base pairs in human DNA. As a consequence, it will soon be possible to easily determine the genetic profile of individuals. 
Genomics and ProteomicsGenomics is the study of the genome, and proteomics is the study of the proteome—all the proteins active in an organism or cell. Proteomics is expected to provide new medicines for genetic disorders.
13.4		Gene Therapy
During gene therapy, a genetic defect is treated by giving the patient a foreign gene. 
	•	Ex vivo therapyCells are removed from the patient, treated, and returned to the patient.
	•	In vivo therapyA foreign gene is given directly to the patient. 
Thinking Scientifically
	1.	Cystic fibrosis occurs in individuals who have two defective copies of the gene for a protein called cystic fibrosis transmembrane regulator (CFTR). Suppose two people have cystic fibrosis, but one is much more severely affected than the other. In addition, a genetic test for cystic fibrosis is positive for one person, but negative for the other. How do you explain these observations?
	2.	Recently, gene therapy trials were performed on ten infants with X-linked severe combined immunodeficiency syndrome (XSCID), also known as “bubble boy disease.” A normal copy of the gene associated with XSCID was inserted into the virus. The virus was then used to transfer the gene into chromosomes in the patients’ cells. The trial was considered a success because the gene was expressed and the children’s immune systems were restored. However, researchers were shocked and disappointed when two children developed leukemia. How might you explain the development of leukemia in these gene therapy patients?  
Testing Yourself
Choose the best answer for each question.
	1.	The major advantage of chorionic villi sampling over amniocentesis is that it
	a. allows karyotyping to be done earlier.
	b. produces karyotypes with better images of chromosomes.
	c. carries a lower risk of spontaneous abortion.
	d.	
produces a sample that is not contaminated by cells from the mother.
	2.	Human genetic disorders due to changes in chromosome number are caused by
	a. radiation.
	b. chemical mutagens.
	c. nondisjunction at meiosis.
	d. fertilization by two sperm cells.
	3.	An example of a chromosomal mutation that involves two nonhomologous chromosomes is
	a. an inversion.	c. a deletion.
	b. a duplication.	d. a translocation.
	4.	Fill in the genotypes a.–i. of the family members in the following pedigree for an autosomal recessive trait. Shaded individuals are affected.
	5.	Fill in the genotypes a.–h. of the family members in the following pedigree for an autosomal dominant trait. Shaded individuals are affected.
	6.	Fill in the genotypes a.–e. of the family members in the following pedigree for an X-linked recessive trait. Shaded individuals are affected.
For questions 7–10, match the pedigree characteristics to those in the key. Answers can be used more than once. Some questions may have more than one answer.
Key:
	a. X-linked recessive
	b. X-linked dominant
	c. autosomal recessive
	d. autosomal dominant
	7.	All daughters and no sons from affected males express the trait.  
	8.	Males express the trait more frequently than females. 
	9.	Affected children always have at least one affected parent. 
	10.	Affected children can have unaffected parents. 
	11.	Disorders that result from the inability to break down a substance are
	a. cystic fibrosis and hemophilia.
	b. Tay-Sachs disease and phenylketonuria.
	c. color blindness and Tay-Sachs disease.
	d. muscular dystrophy and cystic fibrosis.
For questions 12–20, match the descriptions to the conditions in the key. Answers can be used more than once. Some questions may have more than one answer.
Key:
	a. Tay-Sachs disease	f. Huntington disease
	b. cystic fibrosis	g. familial hypercholesterolemia
	c. phenylketonuria	h. color blindness
	d. sickle cell disease	i. Duchenne muscular dystrophy
	e. Marfan syndrome	j. hemophilia
	12.	Autosomal dominant disorder. 
	13.	Disorder caused by incomplete dominance. 
	14.	Neurological disorder that leads to progressive degeneration of brain cells. 
	15.	X-linked recessive disorder in which muscle tissue wastes away. 
	16.	The most common lethal genetic disorder among U.S. Caucasians. 
	17.	X-linked recessive disorder. 
	18.	Results from the lack of the enzyme hex A, resulting in the storage of its substrate in lysosomes. 
	19.	Results from the inability to metabolize phenylalanine. 
	20.	Caused by abnormal hemoglobin, resulting in red blood cells that are not round. 
	21.	The current estimate for the number of genes in the human genome is
	a. 1,000.	c. 500,000.
	b. 30,000.	d. 3 million.
	22.	Adult chromosomes are most easily acquired for study using
	a. amniotic fluid. 	d. a blood sample.
	b. placental cells.	e. a hair sample.
	c. a saliva sample.
	23.	Proteomics is 
	a.	
the application of computer technologies to the study of the genome.
	b.	
the study of the structure, function, and interaction of cellular proteins.
	c.	
the study of the human genome to better understand how genes work.
	d.	all the genes that occur in a cell.
	e.	the study of a person’s complete genotype, or genetic profile.
	24.	A color-blind female and normal male have children. What are the chances that any sons are color-blind? That any daughters are color-blind? 
	25.	Parents who do not have Tay-Sachs disease produce a child who has Tay-Sachs disease. What is the genotype of all persons involved? 
	26.	A 25-year-old man has Huntington disease. Is it possible for him to father a normal son? Explain.
	27.	A girl has hemophilia. What is her genotype? What are the possible genotypes of the parents?  
Go to www.mhhe.com/maderessentials for more quiz questions.
Bioethical Issue
Myriad Genetics, a genomics and genetic testing company, has been awarded a patent on 47 different mutations of the BRCA1 gene. These DNA sequences are used to detect a major breast cancer gene. The patent gives the company the exclusive right to prevent others from using the sequence to develop genetic tests. Opponents of human DNA patents argue that human genes are being treated like commodities. They suggest that reducing humans to a DNA sequence is an assault on our dignity. Proponents argue that the patent system helps improve human health by stimulating biotechnology companies to invest in genetic screening programs. 
Do you agree that human DNA sequences should be protected by patents? If not, do you support the patenting of DNA sequences of other organisms?
Understanding the Terms
amniocentesis197
bioinformatics209
chorionic villi sampling197
chromosomal mutation198
color blindness204
cystic fibrosis202
deletion198
DNA chip205
DNA probe205
Duchenne muscular
dystrophy204
duplication198
familial hypercholesterolemia 
(FH)204
gene therapy210
genetic counseling196
genetic marker205
genetic profile209
genome208
genomics208
hemophilia204
Huntington disease203
inversion198
karyotype196
Marfan syndrome203
pedigree200
phenylketonuria (PKU)202
proteome209
proteomics209
sickle cell disease203
Tay-Sachs disease202
translocation199
Match the terms to these definitions:
a. _______________ Visual display of an individual’s chromosomes arranged by size.
b. _______________ Procedure for sampling cells from the region where the placenta will develop.
c. _______________ Exchange of chromosome segments between two nonhomologous chromosomes.
d. _______________ Chart of a family’s history for a genetic trait.
e. _______________ Study of the structure, function, and interaction of cellular proteins.
f. _______________ Insertion of genetic material into human cells for the treatment of a disorder.

Figure 13.1Genetic counseling.
A genetic counselor uses genetic information about the family to predict the chances a couple will have a child affected by a genetic disorder. If the woman is pregnant, tests can determine whether the child will be born free of genetic disorders. 
Figure 13.4Duplication.
a. When a piece of chromosome 15 is duplicated and inverted, inv dup 15 syndrome results. b. Children with this syndrome have poor muscle tone and autistic characteristics.
Figure 13.7Autosomal recessive pedigree.
The list gives ways to recognize an autosomal recessive disorder. How would you know that the individual at the * is heterozygous?
Figure 13.8Autosomal dominant pedigree.
The list gives ways to recognize an autosomal dominant disorder. How would you know that the individual at the * is heterozygous?
*See page 213.
Figure 13.10Tay-Sachs disease.
Tay-Sachs disease is a lysosomal storage disease; lipid-filled bodies fill the cell because the enzyme needed to metabolize the lipid is missing from lysosomes. The result is impairment of brain and sensory functions.
Figure 13.11Cystic fibrosis.
This person is undergoing antibiotic and percussion therapy for cystic fibrosis. Antibiotic therapy is used to control lung infections of cystic fibrosis patients. The antibiotic can be aerosolized and administered using a nebulizer. A percussion vest loosens mucus in the lungs.
Figure 13.15Muscular dystrophy.
In muscular dystrophy, the calves enlarge because fibrous tissue develops as muscles waste away due to a lack of the protein dystrophin.
Figure 13.18Ultrasound.
Today, an ultrasound exam can produce three-dimensional images that allow physicians to detect serious abnormalities, such as neural tube abnormalities. 
Check Your Progress
	1.	Contrast the use of genetic markers versus DNA probes for the detection of genetic disorders.
	2.	Explain what an ultrasound image can reveal to a doctor.
Answers:1. A genetic marker is an altered DNA sequence that is detectable in individuals with genetic disorders. A DNA probe is a single-stranded piece of DNA that will bind to the mutant DNA sequence responsible for a genetic disorder.2. Ultrasound can determine the baby’s age and size, as well as the presence of serious conditions, such as spina bifida. It can also determine whether there are multiple babies in the womb.

Figure 13.14Familial hypercholesterolemia (FH).
Familial hypercholesterolemia (FH) is incompletely dominant. Heterozygotes have an abnormally high level of plasma cholesterol, and homozygotes have a higher level still.
Figure 13.19Testing the embryo.
Genetic analysis is performed on one cell removed from an 8-celled embryo. If this cell is found to be free of the genetic defect of concern, and the 7-celled embryo is implanted in the uterus, it develops into a newborn with a normal phenotype.
Figure 13.21Studying genomic differences 
between chimpanzees and humans.
Researchers found that by comparing the human genome to that of a chimpanzee they were able to discover genes that cause human diseases when abnormal. They also concluded that the genes for (a) speech, (b) hearing, and (c) smell may have influenced the evolution of humans.
Check Your Progress
	1.	Describe the goals of genomics research.
	2.	Describe the benefits of knowing your genetic profile.
	3.	What do the fields of proteomics and bioinformatics have in common? 
Answers:1. Genomics research attempts to understand how genes direct growth and development in order to control the activities of cells and organisms.2. A genetic profile will tell you whether or not you are at risk for a genetic disorder. It can also indicate which drugs will be most effective for your particular phenotype.3. Although proteomics studies proteins and bioinformatics studies genes, both depend on the computer.
Figure 13.23Ex vivo gene therapy in humans.
Bone marrow stem cells are withdrawn from the body, an RNA retrovirus is used to insert a normal gene into the host genome, and then the cells are returned to the body.
Figure 13.2Testing for chromosomal mutations.
To test the fetus for an alteration in the chromosome number or structure, fetal cells can be acquired by (a) amniocentesis or (b) chorionic villi sampling. c. Karyotyping then will reveal chromosomal mutations. In this case, the karyotype shows that the newborn will have Down syndrome.  
Check Your Progress
	1.	Describe the relationship between amniocentesis and karyotyping.
	2.	
List the two reasons that chorionic villi sampling allows a karyotype to be determined earlier than amniocentesis.
	3.	Explain why karyotypes are created from chromosomes at metaphase of mitosis.
Answers:1. During amniocentesis, a sample of amniotic fluid is collected. Cells from this fluid are cultured and used for karyotyping.2. Chorionic villi sampling can be performed earlier in pregnancy and does not require a cell culture period.3. The chromosomes are the most condensed during metaphase.
Figure 13.5Translocation.
a. When chromosomes 2 and 20 exchange segments, Alagille syndrome results because the translocation disrupts an allele on chromosome 20. b. Distinctive facial features are one of the characteristics of Alagille syndrome.
Check Your Progress
	1.	Contrast a duplication chromosome with a deletion chromosome.
	2.	Contrast a translocation chromosome with an inversion chromosome.
Answers:1. A duplication chromosome contains a repeated segment, while a deletion chromosome is missing a segment.2. A translocation chromosome contains a segment from another, nonhomologous chromosome. An inversion chromosome contains a segment (from the same chromosome) that has been flipped 180°.
Figure 13.6Inversion.
(Left) A segment is inverted in the sister chromatids of one homologue. Notice that, in the red chromosome, edc occurs instead of cde. (Middle) The nonsister chromatids can align only when the inverted sequence forms an internal loop. After crossing-over, a duplication and a deletion can occur. (Right) The inner nonsister chromatid on the left has AB and ab sequences and neither fg nor FG genes. The inner nonsister chromatid on the right has gf and GF sequences and neither AB nor ab genes.
Check Your Progress
	1.	Explain what a genetic counselor can learn from a pedigree.
	2.	
Explain what is meant by “chance has no memory” in pedigree analysis.
	3.	Explain why more males than females express X-linked recessive disorders.
	4.	
Describe the inheritance pattern of a Y-linked trait.
Answers:1. The counselor might be able to determine the inheritance pattern of the disorder and the chances that a child born to the couple will have the disorder.2. The genotype of one child does not influence the genotypes of its siblings.3. Males always express recessive alleles on the X chromosome. There is no possibility for a second allele to mask the recessive allele.4. It would be passed from father to all sons, and it is only expressed in males.
Figure 13.9X-linked recessive pedigree. 
The list gives ways of recognizing an X-linked recessive disorder—in this case, color blindness.
Check Your Progress
	1.	Compare and contrast the physiological bases of Tay-Sachs disease and phenylketonuria.
	2.	Explain why sickle-shaped red blood cells result in a wide array of symptoms in individuals with sickle cell disease.
Answers:1. Both result from the buildup of a substrate that is not broken down properly. The substance is hexosaminidase A for Tay-Sachs disease and phenylalanine for phenylketonuria.2. The cells cannot pass through narrow capillaries as well, so they block them and break down.
Figure 13.12Sickle cell disease.
Persons with sickle cell disease have sickle-shaped red blood cells because of an abnormal hemoglobin molecule.
Figure 13.13Huntington disease.
Huntington disease is characterized by increasingly serious psychomotor and mental disturbances because of a loss of neurons in the brain.
Figure 13.16Use of a genetic marker to test for a genetic mutation. 
a. In this example, DNA from a normal individual has certain restriction enzyme cleavage sites. b. DNA from another individual lacks one of the cleavage sites, and this loss indicates that the person has a mutated gene. In heterozygotes, half of their DNA would have the cleavage site and half would not have it. (In other instances, the gain in a cleavage site could be an indication of a mutation.)
Figure 13.17Use of a DNA chip to test for mutated genes.
This DNA chip contains rows of DNA probes for mutations that indicate the presence of a particular genetic disorder. If single-stranded DNA fragments derived from an individual’s DNA bind to a probe, the individual has the mutation. Heterozygotes would not have as much binding as homozygotes.
Figure 13.20Testing the egg.
Genetic analysis is performed on a polar body removed from an egg. If the egg is free of a genetic defect, it is used for IVF, and the embryo is implanted in the uterus for further development.
Check Your Progress
	1.	List the three ways a doctor can collect fetal cells for genetic testing.
	2.	Explain how an embryo can be tested for a genetic disorder.
	3.	Explain how egg cells can be tested for a genetic disorder.
Answers:1. Amniocentesis, chorionic villi sampling, fetal cells in the mother’s blood.2. When the embryo contains six to eight cells, one cell is removed and used for testing.3. Meiosis results in the production of an egg cell and at least two polar bodies. The polar bodies can be tested for the genetic disorder, and this information can be used to deduce the genotype of the egg cell.
Figure 13.22Bioinformatics.
New computer programs are being developed to make sense out of the raw data generated by genomics and proteomics. Bioinformatics allows researchers to correlate gene activity and protein function, so that the phenotype can be better understood in molecular terms. 
Check Your Progress
	1.	Explain why bone marrow stem cells are preferred for ex vivo gene therapy.
	2.	Explain how ex vivo gene therapy is used to treat cancer.
	3.	Explain how in vivo gene therapy is used to treat cystic fibrosis.
Answers:1. Bone marrow stem cells are capable of dividing after genetic alteration to produce cells carrying the alteration.2. Immune system cells from the patient are removed and genetically altered so that they display tumor antigens. These altered cells are returned to the patient to stimulate the immune system to kill tumor cells.3. The gene needed to cure the disorder is delivered to the respiratory tract in several ways, including via nasal sprays, adenoviruses, or liposomes.
Figure 13.24Gene therapy patients.
a. This patient was treated with the ADA gene to cure severe combined immunodeficiency. b. This patient was treated with the VEGF gene to alleviate poor coronary circulation.
This machine sequences DNA at the rate of 350,000 nucleotides per day.
One cell of an embryo can be removed and tested for genetic mutations. 
A fluorescent probe (complementary DNA) shows the location of a gene.
Figure 13.3Deletion.
a. When chromosome 7 loses an end piece, the result is Williams syndrome. b. These children, although unrelated, have the same appearance, health, and behavioral problems that are characteristic of Williams syndrome.
2500
Figure 13.25Sites of ex vivo and in vivo somatic gene therapy.
Targeted areas (e.g., brain, skin) receive copies of corrected genes by various methods of gene transfer. For many diseases, genetically modified viruses ferry the corrected gene into the body. Many gene therapy treatments are still undergoing trials.