Applications of Genetics

Learning Objectives:
1. Human genetics
• Pedigree Analysis
• Genetic Screening
• Genetic Counseling
• Gene Therapy
• Human Genome Project
2. Plant and animal breeding
• Artificial Selection
• Cloning
3. Recombinant DNA Technology and its
applications
4. DNA fingerprinting and its applications
5. Implications of genetic manipulation: the
potential benefits, hazards and ethical issues.
Human Genetics
1. Pedigree Analysis

Pedigrees = Family Trees

• One of the central tasks of the human
geneticist
• Pedigree analysis is the construction of family
trees
• Family history information is often collected at
major family gatherings
• A pedigree is used to trace inheritance of a
trait over several generations.
Symbols used in pedigree charts:
Inheritance of Haemophilic trait in the Royal Family
Three primary patterns of inheritance:

1.autosomal recessive
2.autosomal dominant
3.sex-linked (X-chromosomal)
 Autosomal Recessive Pedigree
• Recessive: neither parent has the characteristic
phenotype (disease) displayed by the child
• Autosomal: Gene is on one of the autosomes
(Chromosomes 1-22).
• Phenotype only occurs under homozygous condition
 Autosomal Dominant Pedigree

• Dominant: Affected individuals can appear in every

generation
• Autosomal: Gene is on one of the autosomes
(Chromosomes 1-22).
• Phenotype occurs under both homozygous and
heterozygous conditions
 Sex-Linked Pedigree

• X-linked: The trait is preferentially seen in males,

who are homozygous. Females are
heterozygous "carriers"
• Most X-linked traits are recessive.
• Inheritance of red-green color blindness, an X-linked,
recessive trait:
•Color blind male is father of "carrier" daughters and normal sons.
•Carrier daughters have 50% chance to have color blind sons.
•Color blind male x carrier female can produce color blind daughters
•Can you determine a pedigree for Red-Green Colorblindness?
For more information, please refer to the following
website:
Pedigree analysis problem set available at
http://www.biology.arizona.edu/human_bio/pr
oblem_sets/color_blindness/intro.html
 Genetic screening
• Diagnosis of inherited diseases before symptoms occur
by finding abnormalities in the genes or chromosomes.
Some examples:
•sickle cell anaemia
•cystic fibrosis
•phenylketonuria (PKU)
•Down’s Syndrome
Most of these tests performed on cells removed from the
fetus (pre-natal diagnosis) and even from a pre-
implantation embryo.
Ultrasound

•One of the simplest and easiest genetics screening

tests
•Evaluate the physical characteristics of the fetus
which may help predict certain abnormalities
• Measurements of various body parts can be used to
suggest an increased risk of Down Syndrome
•look for congenital defects such leaky heart valves,
and birth defects such as cleft lip or club foot.
•Ultrasound is also used to guide the physician during
amniocentesis and chorionic villus sampling (CVS).
An example of ultrasound image
Amniocentesis

Remove fetal cells from amniotic fluid 14 to 22

weeks after pregnancy.
Culture the cells and look for
•chromosome abnormalities (e.g., the three
number 21 chromosomes of Down syndrome);
•certain enzymatic defects (e.g., an inability to
metabolize galactose, hence milk);
•the sex of the fetus.
Risks:

1. To the mother: slight discomfort, adverse reaction

to medications used, vaginal bleeding or cramping,
and infection.
2. To the foetus: inadvertent puncture is small
3. To the pregnancy: chance to miscarriage very low
(not more than 1/200)
Couples who may wish to consider
amniocentesis include:
1. Women 35 years of age and over
2. Parents who have had a child with Down's
syndrome or other chromosome abnormality.
3. Couples who are known carriers of a
chromosome rearrangement
4. Couples who have had a child with a malformation
of the brain or spinal cord
Chorionic villus sampling (CVS)

Procedure:
• Suck out placental cells by a tube inserted through
the abdomen or vagina.
Advantages:
• No need to perform cell culture
• Can be performed earlier in pregnancy (10-12
weeks)
• If an abortion is to be performed, it is a simpler
process early in pregnancy
Drawbacks:
• More risky than amniocentesis
• Increase the chance of miscarriage (approximately 0.8%)
• Minor complications such as vaginal bleeding or cramping
occur more frequently following CVS than amniocentesis
• Infection
• Transverse limb defects in infants can be resulted
• Involve the absence of the distal structures of the limb
• May result in the vascular system disruption of the limb
• Risk for transverse limb defects following CVS is
approximately 0.03%-0.10%
Couples who may wish to consider CVS include:

• Women 35 years of age and older

• Parents who have had a child with Down's
syndrome or other chromosome abnormality
• Couples who are known carriers of a chromosome
rearrangement
• Couples who have a family history of a genetic
condition for which testing is available
Common genetic diseases diagnosed by
CVS

•Chromosomal abnormalities such as Down’s,

Klinefelter’s and Turner’s syndromes can be
identified by karyotype analysis.
•The cells can be cultured for DNA analysis that
helps diagnosis of
1. Cystic fibrosis
2. Huntington’s chorea
3. Thalassemia
 Genetic Counseling
Genetic counselors are trained persons who help individual
and family to

• comprehend the medical facts, including the diagnosis,

probable course of the disorder, and the available management;
• appreciate the way heredity contributes to the disorder, and the
risk of recurrence in specified relatives
• understand the alternatives for dealing with the risk of
occurrence
• choose the course of action which seems to them appropriate
in view of their risk, their family goals, and their ethical and
religious standards, to act in accordance with that decision
• make the best possible adjustment to the disorder in an
affected family member and/or the risk of recurrence of that
disorder.
Who Should be Referred?

1. Families in which one or more members have a serious

birth defect or genetic disease.
2. Families who have a child with multiple congenital
anomalies, serious developmental delay, or an
unexplained abnormality of growth.
3. Families in which more than one close relative has the
same disease, e.g., mental retardation, deafness,
blindness, cancer, early heart attacks, or schizophrenia.
4. Couples who have had repeated stillbirths or a stillborn
baby with birth defects.
5. Couples who are close blood relatives, e.g., first cousins.
Common issues discussed during genetic
counseling
1. Making a diagnosis
2. Investigate the family history for previous cases of
genetic diseases through pedigree analysis
3. calculate and explain the risk of having affected
children
4. explain the cause of the disease
5. Quality of life
• The quality and likely length of an affected child’s life
will be discussed
• Availability of treatment, support groups and financial
help will also be discussed.
• Effects on other family members will be discussed
 Options:

• These include contraception, sterilization,

adoption, artificial insemination to avoid the
husband’s genes being passed on or IVF using a
donor egg if the problem lies with the woman.
• Prenatal diagnosis is an option if the couple are
willing to consider abortion of an affected fetus.
• CVS, amniocentesis and abortion must be
discussed.
• The reliability of tests such as DNA analysis, ultra-
sound scanning must be explained.
• Possibility of IVF and gene therapy
Ethical issues in genetic counseling

During postnatal counsel

•Risk evaluation, preventive options, gene therapy,
testing of all family members, confidentiality within
the couple
During prenatal counsel
•Option of pregnancy termination, selective
implantation, sex selection
Problems of Eugenics (The birth of designer
baby)
Gene Therapy

A new method for treating genetic diseases by replacing

faulty genes with normal genes or adding normal genes
if they are absent.
Two types of gene therapy:
Somatic cell gene therapy - the insertion of genes
into body cells which function only in the treated person
and are not passed on to the offspring.
Germline gene therapy - the insertion of genes into
cells that are involved in reproduction which can be
passed on to the future offspring.
Diagrammatic Illustration of Gene Therapy
General Procedure of Gene Therapy:
• Isolate and clone the normal gene.
• Introduce it into the chosen human cells with the
help of a safe and efficient vector.
• The cells may have to be isolated from the body
first, corrected and then replaced.
• Easier for blood diseases such as sickle cell
anaemia because the cells that make blood cells
can easily be removed from bone marrow and
replaced.
• The final problem is to make sure that the gene is
expressed normally.
Common Vectors Used:

•Viruses (e.g. Retrovirus) carrying the required gene

•Liposomes containing the cDNA clone
•Microinjection and electroporation- Direct injection of
donor DNA into the cell
Applications of Gene Therapy
•e.g. Cystic fibrosis
•Cystic fibrosis affects the epithelial cells of the body, but the life-
threatening problems mainly affect the lungs.
•Lung and trachea epithelial cells are therefore the initial targets for
gene therapy. The aim is to get the gene into the cells so that it can
make the normal protein, known as CFTR
•The cDNA clone is enclosed in a specially designed vector.
•Adenovirus infecting the respiratory tract is currently used as the
vector for the CFTR gene
•The vector does not insert its DNA into the host DNA. If the cell
divides, the new DNA is not replaced at the same time so it
eventually becomes diluted.
•The treatment may only be effective for a few weeks until the
epithelial cells die, but it will be easy to repeat the treatment at
regular intervals.
Advantages and Disadvantages of Gene Therapy

Advantages Disadvantages

Addition of normal Only somatic cell therapy

allele will alleviate allowed
symptoms
Avoid need for Not a permanent cure as cells
constant medication shed
Extend life Viral vector may give
expectancy inflammation problems

Possibility of DNA entering cells

other than target cells with
unknown effects
1. Ethical issues of gene therapy
• Germline therapy is controversial because it opens
up the whole field of eugenics.
• The same technique can be used to add genes for
desired characteristics. (For example, the American
public has demonstrated a desire for enhanced
growth of normal children with demands for human
growth hormone)
• Germline therapy is controversial because the
change can be passed on to the children of the
treated person and all subsequent generations.
• Do we have the right to alter the genome of the
future generation?
Human Genome Project (HGP)
The goals of the project are to:
• identify all the approximately 30,000 genes in
human DNA
• determine the sequences of the 3 billion
chemical bases that make up human DNA
• store this information in databases
• develop faster, more efficient sequencing
technologies
• develop tools for data analysis, and
• address the ethical, legal, and social issues
(ELSI) that may arise from the project.
 Applications of HGP

1. Molecular Medicine
• Improved diagnosis of disease
• Earlier detection of genetic predispositions to
disease
• Rational drug design
• Gene therapy and control systems for drugs
• Pharmacogenomics "custom drugs"
2. Microbial Genomics

• New energy sources (biofuels)

• Environmental monitoring to detect
pollutants
• Protection from biological and chemical
warfare
• Safe, efficient toxic waste cleanup
• Understanding disease vulnerabilities and
revealing drug targets
3. Risk Assessment

•Assess health damage and risks caused by

radiation exposure, including low-dose exposures
•Assess health damage and risks caused by
exposure to mutagenic chemicals and cancer-
causing toxins
•Reduce the likelihood of heritable mutations
4. Bioarchaeology, Anthropology, Evolution, and
Human Migration

•study evolution through germline mutations in lineages

•study migration of different population groups based on
female genetic inheritance
•study mutations on the Y chromosome to trace lineage
and migration of males
•compare breakpoints in the evolution of mutations with
ages of populations and historical events
5. DNA Forensics (Identification)

•Identify potential suspects whose DNA may match evidence left at

crime scenes
•Exonerate persons wrongly accused of crimes
•Identify crime and catastrophe victims
•Establish paternity and other family relationships
•Identify endangered and protected species as an aid to wildlife officials
(could be used for prosecuting poachers)
•Detect bacteria and other organisms that may pollute air, water, soil,
and food
•Match organ donors with recipients in transplant programs
•Determine pedigree for seed or livestock breeds
•Authenticate consumables such as caviar and wine
6. Agriculture, Livestock Breeding, and
Bioprocessing
•Disease-, insect-, and drought-resistant crops
•Healthier, more productive, disease-resistant farm
animals
•More nutritious produce
•Biopesticides
•Edible vaccines incorporated into food products
(Biopharming)
•New environmental cleanup uses for plants like
tobacco
Ethnical problems
• Fairness in the use of genetic information by
insurers, employers, courts, schools, adoption
agencies, and the military, among others.
• Who should have access to personal genetic
information, and how will it be used?
• Privacy and confidentiality of genetic information.
• Who owns and controls genetic information?
• Psychological impact and stigmatization due to an
individual's genetic differences.
• How does personal genetic information affect an
individual and society's perceptions of that individual?
More information on HGP is available at

Dolan DNA learning center (2002), Gene Almanac,

[Online] Available at
http://www.dnalc.org/resources/resources.html

http://www.web-and-
flow.com/members/efitzger1/genetics/webquest.htm
(The Human Genome WebQuest)
Selective Breeding of Animals and Plants

Aims:
To select desirable traits and remove
undesirable traits of plants and animals through
artificial selection
Two types of breeding involved in artificial
selection:
(1) Inbreeding
(2) Outbreeding
Inbreeding – mating of closely related individuals
Advantages: Create pure line over time
Better adapted to steady
environment.
For example
Parental genotypes: Ffgg x Ffgg

Gametes: Fg , fg Fg , fg
F1: FFgg Ffgg Ffgg ffgg
(pure breeding)
Disadvantages of inbreeding

1. Increasing the chance for recessive genes

to be homozygous
2. Most recessive genes are undesirable
traits
3. Leads to a high frequency of defects
present at birth
4. Reduce the genetic variability vigour and
fertility of a population.
Outbreeding - mating of individuals that are
unrelated

Advantages:
• Progeny (offspring) are heterozygous and the
bad recessive genes are masked by normal
dominant alleles.
• Hybrid vigour – Progeny are tougher, more
fertile and have a greater chance of survival
• Produces variation/heterozygosity; on which
natural selection can act.
Disadvantage:
No more pure line exists
Examples
The cross of larger, non-sweet-tasted tomato with small,
sweet-tasted tomato.
The cross of high-resistant to pest and disease crop
with higher-yield crop.

Parental genotypes: FFgghhIIjj x

FFGGHHiiJJ
Gametes: FghIj FGHiJ
F1: FfGgHhIiJj

Higher chance of heterozygosity in F1

Result in hybrid vigour
 The hybrids are stronger and bigger in size
than their parents

female hybrid male

parent parent
Artificial Insemination for Domestic Animals

Placing semen within the female reproductive

organs through any means other than sexual
intercourse.
Particularly useful with:
•Conditions preventing vaginal ejaculation
•Sperm with low motility (movement)
•Cervical mucus that is hostile to sperm
•Presence of sperm antibodies
Procedure:
Intracervical insemination (ICI) OR
Intrauterine insemination (IUI)
ICI - Fresh ejaculate (semen) is placed
directly into the cervix (opening to the
uterus) by using a syringe and cannula
(long, slender tube).

IUI - "Washed" semen is placed directly into

the uterus via a catheter through the
cervix.
Success Rates

ICI - 2% or higher chance of pregnancy per

cycle

IUI - Higher probability of pregnancy than ICI.

Generally, 5 - 20% chance of pregnancy
per cycle Rates are higher when AI is
accompanied by superovulation using
fertility drugs.
Typical Procedure Protocol:

Pre-procedure:
• Thorough evaluation of both male and female for suspected
causes of infertility
• Determination of drugs to be used (or not) and timing of
treatment cycle
• Monitoring by ultrasound and blood work to evaluate ovarian
response to drugs and reduce potential side effects
• If drugs result in ovarian hyperstimulation syndrome,
treatment cycle will be discontinued immediately.
• In case of no drug use, patient may be required to monitor
own cycle with ovulation predictor kits and other methods
induction of ovulation by HCG , if necessary
 During procedure:

• Male partner must produce a semen sample,

usually in the clinic (may be collected at home
in some cases), or donor semen must be
available
• Female partner is inseminated while lying
down, feet in stirrups, on examination table
• Female may be advised to refrain from
strenuous physical activity for 24 to 48 hours
after insemination
Advantages of using artificial insemination

1. Sperm from one male can be used for many

females;
2. One ejaculate can be used for several
inseminations;
3. Speeds up selective breeding;
4. Saves cost of keeping male and travelling for
mating
5. Less stress than mating;
6. Allows genetic testing
Disadvantages of using artificial insemination

• Danger of inseminating too many females with

one male causing inbreeding;
• With consequent genetic problem;
• Undetected genetic weakness
• Requires expertise;
• Cost of vet.;
• Sperm damage in storage;
• Not successful in some species ;
Diagram showing how sperms are
collected
 Selective breeding of animals – sperm bank
(for domestic animals and human)

1. Sperms are collected and frozen in liquid nitrogen for

further use.
2. Endangered animal species, e.g. giant pandas can be
bred
3. Sperm from one male (with good quality) can be used
for many females
4. Infertility treatment in human
 Selective breeding of animals –embryo
transfer breeding (for domestic animals and
human)
Procedure:
• Injections of hormone to stimulate and ovulation
in the animals that you want to get the embryos
from.
• The donor is inseminated at normal time
• Seven days later, rinsing out the uterus to extract
the embryos and ova (unfertilised, fertilised or
degenerate)
• Isolation of the good embryos using a microscope
and then transfer into the recipient animals e.g.
surrogate.
Applications:
• Domestic animal e.g. cow
1. Produce up to ten or more progeny per year from
their best cows.
2. Profit from the increased sale of quality genetics
without losing the bloodlines.
3. Extend the productive life of some older cows,
incapable of carrying another calf by producing
further progeny through the use of embryo transfer.
4. Conserve the genetics in their herd through the uses
of, embryo freezing for, export, domestic sale or
future transfers on their own farm.
• Infertility treatment in human