PHILIPPINE REHABILITATION FOUNDATION, INC.

College of Sciences –
Department of Physical Therapy and Occupational Therapy

Module 8: DNA Biotechnology

Module Objectives:
1. To understand ways by which DNA are analyzed
2. To understand gene mapping, testing and the identification and isolation of genes responsible
for genetic disorders

We inherit our physical appearance from a long and complicated lineage that tells the story of who we
are and where we came from. Inheritance of all genes follows the principles of Mendelian genetics. It
wasn't until certain genes began to disobey Mendel's rules, that the concept of linked genes was
discovered.

Without the knowledge of chromosomes, Gregor Mendel laid the groundwork for understanding
inheritance of genes. In the previous modules, we have discussed that genes are found on
chromosomes, and homologous chromosomes segregate independently of one another during meiosis.
Two different genes should end up in sex cells after meiosis in predicted ratios. However, sometimes
certain genes end up together in higher frequencies. This phenomenon occurs when genes are close
together, and on the same chromosome. Genes that are on the same chromosome are defined as linked
(Figure 1).

Figure 1. All 23 pairs of chromosomes in humans. The dark bands represent different genes. Genes on the same
chromosome are linked.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Genetic Linkage

Linked genes travel together, and therefore end up together in the same sex cells after meiosis.
Sometimes genes traveling on the same chromosome can end up switching to another homologous
chromosome. This occurs due to the process of crossing over, and complicates linked inheritance.
Crossing over results in recombination. Recombination means that the offspring have different
combination of traits than their parents. Two genes on the same chromosome that have experienced
crossing over will follow the laws of independent assortment again. It then must be stated that linked
genes located closely together on chromosomes travel together. Linked genes farther apart on
chromosomes may be subject to independently assort due to the phenomenon of crossing over (Figure
2).

Figure 2. While genes AB are initially linked together, after crossing over the two undergo recombination and will be
inherited independently.

Recombinant and Nonrecombinant Offspring

Recombination of traits sets children apart from their parents. There will never be, nor has there ever
been, another you. While the genes that make you do come from your parents, the traits are inherited
in different combinations than your folks. Having new combinations of alleles in gametes is
called recombinant gametes. Crossing over, and recombination of traits from two parents, leads to the
many combinations of traits, making offspring unique from their parents. If progeny contains the same
combination of alleles as the parent, it is called nonrecombinant.

Genes linked closely will not be mixed up, so they will be inherited exactly like the parent, and be
nonrecombinant. By knowing the frequency of appearance of nonrecombinant and recombinant traits in
offspring, the relative distance between genes on a chromosome can be calculated. Thomas Morgan
first realized this in the fruit fly and mapped the genes on their chromosome II. Knowing the distance
apart genes are on a chromosome allows scientist to sketch where genes are located, this is called gene
mapping.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

When genes are on separate chromosomes, or very far apart on the same chromosomes, they assort
independently. That is, when the genes go into gametes, the allele received for one gene doesn't affect
the allele received for the other. In a double heterozygous organism (AaBb), this results in the formation
of all 4 possible types of gametes with equal, or 25% frequency.

If genes separate with independent assortment you would expect to find a predicted ratio of offspring.
If, say, an F1 dihybrid cross is bred with a double mutant, the results should be equal numbers of four
different offspring.
The cross of independent assortment:
(F1 dihybrid) GgLl x ggll (double mutant)
 Gray bodied (G) Black bodied (g) Long wing (L) short wing (l)
Offspring are 1GgLl: 1ggll: 1Ggll 1: ggLl.
Two offspring have the same identical genetics as the parents (GgLl and ggll). Two have unique
combinations (Ggll and ggLl.). In other words, half the offspring are nonrecombinant and the other half
is recombinant. If the genes are very closely linked, and located close together, the ratios will not follow
this test, but instead have all nonrecombinant offspring.

The cross if genes are very close together:

GgLl x ggll
Offspring are all GgLl or ggll

If genes are located far apart on the same chromosome they are more likely to undergo crossing over.
When crossing over occurs, the result is offspring ratios closer to ratios with independent assortment.
The closer offspring are to predicted ratios with independent assortment, the further the genes are on a
chromosome. By knowing the frequency of recombination in offspring, you can calculate the distance
between genes on a chromosome.

Distance Between Genes

Thomas Morgan's early experiments showed that frequency of recombination of offspring not only
shows if genes are linked, but the distance between those genes on the chromosome. In his test he
crossed a heterozygous fly, for the traits of body color and wing shape, with a double mutant fly. The
heterozygous fly had the wild type traits of gray body and normal wings, GgLl. The double mutant had
black body and small wings, ggll. The resultant 2,300 offspring were:

 965 GgLl
 944 ggll
 206 Ggll
 185 ggLl

If you add the recombinant offspring (Ggll & ggLl), 391 offspring are unique form the parents (Figure 3).
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Figure 3. A testcross of two genes to see if they are linked. 2,300 offspring are produced and the results show more
nonrecombinant offspring. The results provide evidence that genes are linked. If genes were on different
chromosomes, they would show 1:1:1:1 ratios

The ratio is not close to a 1:1:1:1, and not all offspring are nonrecombinant. Therefore, the frequency of
recombinant offspring is 391/2,300=.17 or 17 percent. This tells us that the distance between the genes
is far enough apart that 17 percent of the time, crossing over occurs and allows for recombinant
offspring. This first part explained that when genes are linked, offspring have a higher frequency
of nonrecombinant offspring. Recombination occurs due to crossing over and independent assortment,
and leads to recombinant offspring, which are not identical to the parents. By comparing the frequency
of recombinant offspring, you can determine how far apart two genes are on a chromosome, a process
called gene mapping.

What is the benefit of calculating recombination frequency? One way that recombination frequencies
have been used historically is to build linkage maps, chromosomal maps based on recombination
frequencies. In fact, studying linkage helped early geneticists establish that chromosomes were in fact
linear, and that each gene had its own specific place on a chromosome. By doing this type of analysis
with more and more genes (e.g., adding in genes D, E, and F and figuring out their relationships to A, B,
and C) we can build up linkage maps of entire chromosomes. In linkage maps, you may see distances
expressed as centimorgans or map units rather than recombination frequencies. Luckily, there's a direct
relationship among these values: a 1% recombination frequency is equivalent
to 1 centimorgan or 1 map unit.

Map distance is sometimes the same as recombination frequency. Sometimes, the directly measured
recombination frequency between two genes is not the most accurate measure of their map distance.
That's because, in addition to the single crossovers, double crossovers (two separate crossovers
between the two genes) can also occur.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Gene Mapping

Gene mapping is assigning/locating of a specific gene to particular region of a chromosome and

determining the location of and relative distances between genes on the chromosome. Gene maps help
describe the spatial arrangement of genes on a chromosome. Genes are designated to a specific location
on a chromosome known as the locus and can be used as molecular markers to find the distance
between other genes on a chromosome. Maps provide researchers with the opportunity to predict the
inheritance patterns of specific traits, which can eventually lead to a better understanding of disease-
linked traits.

The genetic basis to gene maps is to provide an outline that can potentially help researchers carry
out DNA sequencing. A gene map helps point out the relative positions of genes and allows researchers
to locate regions of interest in the genome. Genes can then be identified quickly and sequenced quickly.
Two approaches to generating gene maps include physical mapping and genetic mapping. Physical
mapping utilizes molecular biology techniques to inspect chromosomes. These techniques consequently
allow researchers to observe chromosomes directly so that a map may be constructed with relative
gene positions. Genetic mapping on the other hand uses genetic techniques to indirectly find association
between genes. Techniques can include cross-breeding experiments and examining pedigrees. These
technique allow for maps to be constructed so that relative positions of genes and other important
sequences can be analyzed (Figure 4).

Figure 4. Types of gene mapping

Physical maps. Represent chromosomes and provide physical distances between chromosomal
landmarks ideally measured in nucleotide bases. There are different types of physical maps, as described
below but the ultimate physical map is the complete sequence itself.

Physical mapping techniques used to generate a gene map include: Restriction mapping, Fluorescent in
situ hybridization (FISH), and Sequence tagged site (STS) mapping.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

 Restriction Mapping. Restriction mapping is a method in which structural information regarding

a segment of DNA is obtained using restriction enzymes. Restriction enzymes are enzymes that
help cut segments of DNA at specific recognition sequences. The basis to restriction mapping
involves digesting (or cutting) DNA with restriction enzymes. The digested DNA fragments are
then run on an agarose gel using electrophoresis, which provides one with information
regarding the size of these digested fragments. The sizes of these fragments help indicate the
distance between restriction enzyme sites on the DNA analyzed, and provides researchers with
information regarding the structure of DNA analyzed.
 Fluorescent in situ Hybridization (FISH). FISH is a method used to detect the presence (or
absence) of a DNA sequence within a cell. DNA probes that are specific for chromosomal regions
or genes of interest are labeled with fluorochromes. By attaching fluorochromes to probes,
researchers are able to visualize multiple DNA sequences simultaneously. When a probe comes
into contact with DNA on a specific chromosome, hybridization will occur. Consequently,
information regarding the location of that sequence of DNA will be attained. FISH analyzes single
stranded DNA (ssDNA). Once the DNA is in its single stranded state, the DNA can bind to its
specific probe.
 Sequence Tagged Site (STS) Mapping. A sequence tagged site (STS) is a short sequence of DNA
(about 100 - 500 base pairs in length) that is seen to appear multiple times within an individual's
genome. These sites are easily recognizable, usually appearing at least once in the DNA being
analyzed. These sites usually contain genetic polymorphisms making them sources of viable
genetic markers (as they differ from other sequences). Sequenced tagged sites can be mapped
within our genome and require a group of overlapping DNA fragments. PCR is generally used to
produce the collection of DNA fragments. After overlapping fragments are created, the map
distance between STSs can be analyzed. In order to calculate the map distance between STSs,
researchers determine the frequency at which breaks between the two markers occur
(see shotgun sequencing).

Genetic Maps. Genetic mapping, also called linkage mapping show the arrangement of genes and
genetic markers along the chromosomes as calculated by the frequency with which they are inherited
together (Figure 5). Among the main goals of the Human Genome Project (HGP) was to develop new,
better and cheaper tools to identify new genes and to understand their function. One of these tools is
genetic mapping. It can offer firm evidence that a disease transmitted from parent to child is linked to
one or more genes. Mapping also provides clues about which chromosome contains the gene and
precisely where the gene lies on that chromosome. Genetic maps have been used successfully to find
the gene responsible for relatively rare, single-gene inherited disorders such as cystic fibrosis and
Duchenne muscular dystrophy. Genetic maps are also useful in guiding scientists to the many genes that
are believed to play a role in the development of more common disorders such as asthma, heart
disease, diabetes, cancer, and psychiatric conditions.

To produce a genetic map, researchers collect blood or tissue samples from members of families in
which a certain disease or trait is prevalent. Using various laboratory techniques, the scientists isolate
DNA from these samples and examine it for unique patterns that are seen only in family members who
have the disease or trait. These characteristic patterns in the chemical bases that make up DNA are
referred to as markers.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Markers themselves usually consist of DNA that does not contain a gene. But because markers can help
a researcher locate a disease-causing gene, they are extremely valuable for tracking inheritance of traits
through generations of a family.

DNA markers don't, by themselves, identify the gene responsible for the disease or trait; but they can
tell researchers roughly where the gene is on the chromosome. Because of recombination, the single
chromosome in a reproductive cell contains some stretches of DNA inherited from the person's mother
and some from his or her father. If a particular gene is close to a DNA marker, the gene and marker will
likely stay together during the recombination process, and they will likely be passed on together from
parent to child. If each family member with a particular disease or trait also inherits a particular DNA
marker, it is very likely that the gene responsible for the disease lies near that marker. The more DNA
markers there are on a genetic map, the more likely it is that at least one marker will be located close to
a disease gene-and the easier it will be for researchers to zero in on that gene. One of the first major
achievements of the HGP was to develop dense maps of markers spaced evenly across the entire human
genome. Genetic mapping is focused on the principles first established by Gregor Mendel. This approach
primarily focuses on linkage analysis and gene association techniques.
 Linkage Analysis. The basis to linkage analysis is understanding chromosomal location and
identifying disease genes. Certain genes that are linked or associated with each other are found
to reside close to each other on the same chromosome. During meiosis, these genes are capable
of being inherited together and can be used as a genetic marker to help identify
the phenotype of diseases. Because linkage analysis can identify inheritance patterns, these
studies are usually family based.
 Gene Association Analysis. Gene association analysis is population based; it is not focused on
inheritance patterns, but rather is based on the entire history of a population. Gene association
analysis looks at a particular population and tries to identify whether the frequency of
an allele in affected individuals is different from that of a control set of unaffected individuals of
the same population. This method is particularly useful to identify complex diseases that do not
have a Mendelian inheritance pattern.

Figure 5. A gene map of Drosophila melanogaster, showing the relative positions of genes on one of Drosophila’s
chromosomes.

The development of easy-to-use genetic maps, coupled with the HGP's successful sequencing of the
entire human genome, has greatly advanced genetics research. The improved quality of genetic data has
reduced the time required to identify a gene from a period of years to, in many cases, a matter of
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

months or even weeks. Genetic mapping data generated by the HGP's laboratories is freely accessible to
scientists through databases maintained by the National Institutes of Health and the National Library of
Medicine's National Center for Biotechnology Information (NCBI) [ncbi.nlm.nih.gov], as well as
the Genome Browser of University of California, Santa Cruz.

Biotechnology

The term biotechnology refers to any technology, process or practice that modifies or harnesses any
living organism or system to be useful to any human purpose. The word defines itself: bio means life
and technology is defined as the application, or harnessing of science for a specific purpose.

Brief History of Biotechnology

It is not known precisely when people began using the word biotechnology to refer to technologies
based on living things. Many believe the phrase was first coined in the early 1900s by a Hungarian
engineer named Károly Ereky, but the practice of biotechnology began long before that and dates back
almost to the beginning of human existence. The following are examples of biotechnologies over the
course of history:
 Agriculture, the practice of purposefully growing plants, which are living organisms, for
consumption or other use, is one of the earliest forms of biotechnology. When ancient humans
began to transition from the hunter-gatherer lifestyle and settled into societies, they were able
to do so by cultivating food in one place. Agricultural technology has itself evolved immensely
over thousands of years, and all advancements from crop rotation, beginning in ancient Greece,
to the creation of genetically modified organisms, or GMOs, today relate to this type of
biotechnology.
 Animal husbandry, the process of housing, feeding, breeding and utilizing animals, is as much a
pillar of human existence as agriculture. When humans domesticated animals to be beasts of
burden such as oxen, modes of transportation such as horses, or even companions such as the
dog, biotechnology was implemented.
 Fermentation of beer and leavening of bread, two ancient and widespread methods of food
preparation, are examples of early biotechnologies. Both processes require the use of the living
organism yeast, which the ancient Chinese (7 BCE) and Egyptians (4 BCE) first utilized for these
processes, respectively.
 Vaccinations are one of the earliest examples of biotechnology applied to
medicine. Vaccinations are the administration of a deactivated version of a virus that normally
causes an illness into a healthy person. This teaches the immune system to look out for that
virus in the future, and the vaccinated person becomes immune to the illness as a result.
Although there is debate over whether a virus is truly 'alive,' vaccinations fall squarely under the
umbrella of biotechnology. The first vaccination was developed at the turn of the 19th century
to protect healthy people from smallpox.
 Antibiotics are drugs that combat the growth of bacteria and are used to treat common illnesses
such as strep throat and sinus infections. The first antibiotic was devised in the early 1900s
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

when a species of mold was discovered to halt the expansion of bacteria. Penicillin was later
isolated from the mold and is still used frequently today.
Modern Examples of Biotechnology
Biotechnology has exploded in the late 20th century with the ability to sequence and manipulate
genomes. The following are some noteworthy areas of expansion:
 Genetic testing, the process of examining an individual's genome to determine predispositions
to certain diseases, is made possible by our ability to read DNA and our understanding of the
entire human genetic code, which was made possible by the completion of the Human Genome
Project in 2000.
 Cloning, the controversial process of creating a new organism that is identical to an existing
organism, has been in the news since Dolly the sheep was cloned in the 1990s.
 Genetically modified (GM) foods were first approved by the US FDA in 1994. The earliest
example was a tomato that had longer lasting flavor.
 Biotechnology drugs are generated by bacteria with modified genomes. For example, in the
1980s, bacteria were mutated to reprogram them to generate insulin for treating diabetes.
Biotechnology also refers to technological advancements that interface with living things for a human
purpose. The medical field of prosthetic development, for example, is a biotechnology. The most
technologically advanced prosthetics communicate directly with the nervous system and can be
controlled by the patient's brain as if they were part of the body.

DNA Technology

Many forms of modern biotechnology rely on DNA technology. DNA technology is the sequencing,
analysis, and cutting-and-pasting of DNA. Many examples of modern biotechnology depend on the
ability to analyze, manipulate, and cut and paste pieces of DNA. Approaches for the sequencing and
manipulation of DNA are sometimes referred to as DNA technology. For example, for the cystic fibrosis
gene therapy trial, researchers used DNA manipulation techniques to insert the chloride channel gene
into a piece of carrier DNA (a vector) that allowed it to be expressed in human lung cells.
DNA technology is important to both basic and applied (practical) biology. For instance, a technique
used to make many copies of a DNA sequence, called polymerase chain reaction (PCR), is used in many
medical diagnostic tests and forensics applications as well as in basic laboratory research.

Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning,
and gel electrophoresis.

 DNA cloning. In DNA cloning, researchers “clone” – make many copies of – a DNA fragment of
interest, such as a gene. In many cases, DNA cloning involves inserting a target gene into a
circular DNA molecule called a plasmid. The plasmid can be replicated in bacteria, making many
copies of the gene of interest. In some cases, the gene is also expressed in the bacteria, making
a protein (such as the insulin used by diabetics) (Figure 6).
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Figure 6. Insertion of a gene into a plasmid. A plasmid is a small DNA molecule within a cell that is physically
separated from a chromosomal DNA and can replicate independently. They are most commonly found as small
circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and
eukaryotic organisms.

 Polymerase chain reaction (PCR). Polymerase chain reaction is another widely used DNA
manipulation technique, one with applications in almost every area of modern biology. PCR
reactions produce many copies of a target DNA sequence starting from a piece of template DNA.
This technique can be used to make many copies of DNA that is present in trace amounts (e.g.,
in a droplet of blood at a crime scene).

 Gel electrophoresis. Gel electrophoresis is a technique used to visualize DNA fragments. For
instance, researchers can analyze the results of a PCR reaction by examining the DNA fragments
it produces on a gel. Gel electrophoresis separates DNA fragments based on their size, and the
fragments are stained with a dye so the researcher can see them (Figure 7).

Figure 7. DNA fragments migrate through the gel from the negative to
the positive electrode.

After the gel has run, the fragments are separated by size, with the
smallest ones near the bottom (positive electrode) and the largest ones
near the top (negative electrode).
(bp = base pairs)
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

 DNA sequencing. DNA sequencing involves determining the sequence of nucleotide bases (As,
Ts, Cs, and Gs) in a DNA molecule. In some cases, just one piece of DNA is sequenced at a time,
while in other cases, a large collection of DNA fragments (such as those from an entire genome)
may be sequenced as a group.

Diagnosing and Screening for Genetic Disorders

Genetic testing is "the analysis of chromosomes, DNA, proteins, and certain metabolites in order to
detect heritable disease-related genotypes, mutations, phenotypes, or karyotypes for clinical
purposes.” It can provide information about a person's genes and chromosomes throughout life. The
variety of genetic tests has expanded throughout the years. In the past, the main genetic tests searched
for abnormal chromosome numbers and mutations that lead to rare, inherited disorders. Today, tests
involve analyzing multiple genes to determine the risk of developing specific diseases or disorders, with
the more common diseases consisting of heart disease and cancer. The results of a genetic test can
confirm or rule out a suspected genetic condition or help determine a person's chance of developing or
passing on a genetic disorder. Several hundred genetic tests are currently in use, and more are being
developed.

Much diagnosis for genetic diseases is performed prenatally. The most widely used methods for prenatal
diagnosis of genetic disorders are amniocentesis and chorionic villus sampling (CVS). In amniocentesis, a
needle is used to withdraw amniotic fluid (Figure 8). The fluid and the cells it contains can be analyzed
for chromosomal or single-gene disorders. In CVS, a catheter is inserted into the uterus and used to
retrieve a small tissue sample of the fetal chorion. This tissue is used for cytogenetic, biochemical and
recombinant DNA-based testing.

Figure 8. Amniocentesis. The position of the fetus is first determined by ultrasound, and then a needle is inserted
through the abdominal and uterine walls to recover fluid and fetal cells for cytogenetic and/or biochemical analysis.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

Types of genetic testing include:

 Cell-free fetal DNA (cffDNA) testing is a non-invasive (for the fetus) test. It is performed on a
sample of venous blood from the mother, and can provide information about the fetus early in
pregnancy. As of 2015 it is the most sensitive and specific screening test for Down syndrome.

 Newborn screening: Newborn screening is used just after birth to identify genetic disorders that
can be treated early in life. A blood sample is collected with a heel prick from the newborn 24 –
48 hours after birth and sent to the lab for analysis. All states in the US, by law test for at least
21 disorders. If abnormal results are obtained, it does not necessarily mean the child has the
disorder. Diagnostic tests must follow the initial screening to confirm the disease. All states
currently test infants for phenylketonuria (a genetic disorder that causes mental illness if left
untreated) and congenital hypothyroidism (a disorder of the thyroid gland). Newborn screening
can detect the presence of PKU, allowing kids to get put on a special diet right away to avoid the
effects of the disorder. The aim of newborn screening is to identify conditions that are treatable
in order to begin treatment as soon as possible to prevent serious mental or physical handicaps.

 Diagnostic testing: Diagnostic testing is used to diagnose or rule out a specific genetic or
chromosomal condition. In many cases, genetic testing is used to confirm a diagnosis when a
particular condition is suspected based on physical mutations and symptoms. Diagnostic testing
can be performed at any time during a person's life, but is not available for all genes or all
genetic conditions. The results of a diagnostic test can influence a person's choices about health
care and the management of the disease. For example, people with a family history of polycystic
kidney disease (PKD) who experience pain or tenderness in their abdomen, blood in their urine,
frequent urination, pain in the sides, a urinary tract infection or kidney stones may decide to
have their genes tested and the result could confirm the diagnosis of PKD.

 Carrier testing: Carrier testing is used to identify people who carry one copy of a gene mutation
that, when present in two copies, causes a genetic disorder. This type of testing is offered to
individuals who have a family history of a genetic disorder and to people in ethnic groups with
an increased risk of specific genetic conditions. Examples of Carrier Screening:
o Tay Sachs disease in Ashkenazi Jewish populations (1/27)
o Sickle cell anemia in African American populations (1/13)
o Cystic fibrosis in Caucasian populations (1/25)

 Preimplantation genetic diagnosis: Genetic testing procedures that are performed on human
embryos prior to the implantation as part of an in vitro fertilization procedure. Pre-implantation
testing is used when individuals try to conceive a child through in vitro fertilization. Eggs from
the woman and sperm from the man are removed and fertilized outside the body to create
multiple embryos. The embryos are individually screened for abnormalities, and the ones
without abnormalities are implanted in the uterus.

 Prenatal diagnosis: Used to detect changes in a fetus's genes or chromosomes before birth. This
type of testing is offered to couples with an increased risk of having a baby with a genetic or
chromosomal disorder. In some cases, prenatal testing can lessen a couple's uncertainty or help
them decide whether to abort the pregnancy. It cannot identify all possible inherited disorders
and birth defects, however. One method of performing a prenatal genetic test involves an
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

amniocentesis as mentioned above. The fluid may be tested for chromosomal abnormalities
such as Down syndrome (Trisomy 21) and Trisomy 18, which can result in neonatal or fetal
death. Test results can be retrieved within 7–14 days after the test is done. This method is
99.4% accurate at detecting and diagnosing fetal chromosome abnormalities. Although there is
a risk of miscarriage associated with an amniocentesis, the miscarriage rate is only 1/400.
Another method of prenatal testing is Chorionic Villus Sampling (CVS). Chorionic villi are
projections from the placenta that carry the same genetic makeup as the baby. During this
method of prenatal testing, a sample of chorionic villi is removed from the placenta to be
tested. This test is performed 10–13 weeks into pregnancy and results are ready 7–14 days after
the test was done. Another test using blood taken from the fetal umbilical cord is percutaneous
umbilical cord blood sampling. The aim of prenatal testing is to enable parents to have children
they otherwise would not have been willing to have because of a fear of birth defects or genetic
disease.

 Predictive and presymptomatic testing: Predictive and presymptomatic types of testing are
used to detect gene mutations associated with disorders that appear after birth, often later in
life. These tests can be helpful to people who have a family member with a genetic disorder, but
who have no features of the disorder themselves at the time of testing. Predictive testing can
identify mutations that increase a person's chances of developing disorders with a genetic basis,
such as certain types of cancer. For example, an individual with a mutation in BRCA1 has a 65%
cumulative risk of breast cancer. Hereditary breast cancer along with ovarian cancer syndrome
are caused by gene alterations in the genes BRCA1 and BRCA2. Major cancer types related to
mutations in these genes are female breast cancer, ovarian, prostate, pancreatic, and male
breast cancer. Li-Fraumeni syndrome is caused by a gene alteration on the gene TP53. Cancer
types associated with a mutation on this gene include breast cancer, soft tissue sarcoma,
osteosarcoma (bone cancer), leukemia and brain tumors. In the Cowden syndrome there is a
mutation on the PTEN gene, causing potential breast, thyroid or endometrial
cancer. Presymptomatic testing can determine whether a person will develop a genetic
disorder, such as hemochromatosis (an iron overload disorder), before any signs or symptoms
appear. The results of predictive and presymptomatic testing can provide information about a
person’s risk of developing a specific disorder, help with making decisions about medical care
and provide a better prognosis.

 Pharmacogenomics: type of genetic testing that determines the influence of genetic variation
on drug response. When a person has a disease or health condition, pharmacogenomics can
examine an individual’s genetic makeup to determine what medicine and what dosage would be
the safest and most beneficial to the patient. In the human population, there are approximately
11 million single nucleotide polymorphisms (SNPs) in people’s genomes, making them the most
common variations in the human genome. SNPs reveal information about an individual’s
response to certain drugs. This type of genetic testing can be used for cancer patients
undergoing chemotherapy. A sample of the cancer tissue can be sent in for genetic analysis by a
specialized lab. After analysis, information retrieved can identify mutations in the tumor which
can be used to determine the best treatment option.

Genetic testing is often done as part of a genetic consultation and as of mid-2008 there were more than
1,200 clinically applicable genetic tests available. Once a person decides to proceed with genetic testing,
a medical geneticist, genetic counselor, primary care doctor, or specialist can order the test after
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

obtaining informed consent. Genetic tests are performed on a sample of blood, hair, skin, amniotic
fluid (the fluid that surrounds a fetus during pregnancy), or other tissue. For example, a medical
procedure called a buccal smear uses a small brush or cotton swab to collect a sample of cells from the
inside surface of the cheek. Alternatively, a small amount of saline mouthwash may be swished in the
mouth to collect the cells. The sample is sent to a laboratory where technicians look for specific changes
in chromosomes, DNA, or proteins, depending on the suspected disorders, often using DNA sequencing.
The laboratory reports the test results in writing to a person's doctor or genetic counselor.
Routine newborn screening tests are done on a small blood sample obtained by pricking the baby's heel
with a lancet.

The physical risks associated with most genetic tests are very small, particularly for those tests that
require only a blood sample or buccal smear (a procedure that samples cells from the inside surface of
the cheek). The procedures used for prenatal testing carry a small but non-negligible risk of losing the
pregnancy (miscarriage) because they require a sample of amniotic fluid or tissue from around the fetus.
Many of the risks associated with genetic testing involve the emotional, social, or financial
consequences of the test results. People may feel angry, depressed, anxious, or guilty about their
results. The potential negative impact of genetic testing has led to an increasing recognition of a "right
not to know". In some cases, genetic testing creates tension within a family because the results can
reveal information about other family members in addition to the person who is tested. The possibility
of genetic discrimination in employment or insurance is also a concern. Some individuals avoid genetic
testing out of fear it will affect their ability to purchase insurance or find a job. Health insurers do not
currently require applicants for coverage to undergo genetic testing, and when insurers encounter
genetic information, it is subject to the same confidentiality protections as any other sensitive health
information. In the United States, the use of genetic information is governed by the Genetic Information
Nondiscrimination Act (GINA).

Genetic testing can provide only limited information about an inherited condition. The test often can't
determine if a person will show symptoms of a disorder, how severe the symptoms will be, or whether
the disorder will progress over time. Another major limitation is the lack of treatment strategies for
many genetic disorders once they are diagnosed.
Another limitation to genetic testing for a hereditary linked cancer, is the variants of unknown clinical
significance. Because the human genome has over 22,000 genes, there are 3.5 million variants in the
average person's genome. These variants of unknown clinical significance means there is a change in the
DNA sequence, however the increase for cancer is unclear because it is unknown if the change affects
the gene's function.

A genetics professional can explain in detail the benefits, risks, and limitations of a particular test. It is
important that any person who is considering genetic testing understand and weigh these factors before
making a decision. Other risks include accidental findings—a discovery of some possible problem found
while looking for something else.

The Ethics of Genetic Testing

Gene testing, although helpful in identifying those at risk for incurring a genetic disorder and finding out
the prognosis of an individual’s medical condition, the use of this technology also raises legal, social and
ethical issues which are not yet completely resolved. Although we have the ability to test for many
genetic diseases, there are almost no effective treatments to cure or improve most of these disorders. It
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

is important to remember that with present technology, the fact that a genetic test gives a negative
result does not rule out future development of the disease, nor does a positive result always mean that
one will get the disease. The Ethical, Legal and Social Implications (ELSI) Program of the Human Genome
Project has set up task forces to identify issues related to genetic testing, This program is charged with
developing recommendations for policy makers and lawmakers that will retain the benefits of genetic
testing, but reduce or eliminate the potential harm. In addition, other groups made up of scientists,
health care professionals, lawmakers, ethicists, and consumers are debating these issues and
formulating policy options.

Gene Therapy

Gene products, such as insulin, have been used for decades in therapeutic treatment. Methods for the
isolation and cloning of specific genes, originally developed as a research tool, are now being used to
treat genetic disorders, a process known as gene therapy. In theory, gene therapy transfers a normal
allele into a somatic cell that carries one or more mutant alleles. The delivery of these structural genes
anf their regulatory sequences is accomplished using a vector or gene transfer system. Several methods
can be used to transfer genes and their regulatory sequences into human cells. These include the use of
viruses as vectors, the chemically assisted transfer of genes across cell membranes, and the fusion of
cells with artificial vesicles containing cloned DNA sequences.

Several heritable disorders are currently being treated with gene therapy, including severe combined
immunodeficiency disorder (SCID), familial hypercholesterolemia, and cystic fibrosis. In SCID, affected
individuals have no functional immune system and usually die from otherwise minor infections. An
autosomal form of SCID is caused by a mutation in the gene that encodes for the enzyme adenosine
deaminase (ADA). Treatment starts with the isolation from the patient of a subpopulation of white
blood cells called T cells. These T cells, which are part of the immune system, are mixed with a
genetically modified retrovirus carrying a normal copy of the human ADA gene. The virus infects the T
cells, inserting a functional copy of the ADA gene into the cell’s genome. The genetically modified T cells
are grown in the laboratory to ensure that the transferred gene is expressed, and the patient is treated
by injecting a billion or so of the altered T cells into the bloodstream.

Gene therapy began in 1990, with the treatment of a young girl suffering from SCID. Three years after
treatment, she had a new ADA gene in more than 50% of her T cells, and she is leading a normal life. A
second child, treated a short time later, had the normal gene in only 0.1 to 1% of her white blood cells, a
level that was not high enough to be effective. Other gene therapy trials, such as those for cystic
fibrosis, have also had poor results, most of which have been traced to inefficient vectors.

In addition to treating heritable disorders, gene therapy is now being used or contemplated as a
treatment for skin cancer, breast cancer, brain cancer and AIDS. Treatment of these conditions by gene
therapy has been more successful than that of metabolic disorders.
 PHILIPPINE REHABILITATION FOUNDATION, INC. College of Sciences –
Department of Physical Therapy and Occupational Therapy

References:

 Klug WS, Cummings MR, Spencer CA and Palladino MA. Essentials of Genetics, 9th Ed., 2015.*
 https://www.khanacademy.org/science/biology/biotech-dna-technology/intro-to-biotech-
tutorial/a/intro-to-biotechnology
 https://study.com/academy/lesson/what-is-biotechnology-definition-history-examples.html
 https://www.ncbi.nlm.nih.gov/probe/docs/applmapping/
 https://www.genome.gov/10000715/genetic-mapping-fact-sheet/
 https://en.wikipedia.org/wiki/Gene_map
 https://en.wikipedia.org/wiki/Genetic_testing