Medical

Biotechnology
Chapter 11
Testing for Down’s Syndrome and sex

“Karyotyping”
 Screening for genetic abnormalities
• Fluorescent in situ
hybridization (FISH)
used to detect:
– Extra chromosomes
– Missing parts of
chromosomes
– DNA swapping
across different
chromosomes
• Chronic
myelogenous
leukemia
– DNA exchange ACCATG GTATAC
between
chromosome 9
and 22
*TGGTAC *CATATG
Fluorescent DNA probes
 Allele specific oligonucleotide analysis (ASO)
• Analyze DNA from
cells of 8-32-cell-
stage-old embryo
created by in vitro
fertilization
• Allows individuals
to select health
embryos before
implantation
 Single nucleotide polymorphisms

Oligonucleotide probes that

react with sequence of
normal A gene or disease S
gene
Cell DNA

GAGGACTCCTCTTCA
Extract CTCCTGAGGAGAAGT
DNA
nucleus pcr amplify
primer
GAGGACTCCTCTTCA
CTCCTGAGGAGAAGT
primer

Add probe for dot blot

normal gene Add probe for
disease gene

Normal gene sequence Disease gene sequence

GAGGACTCCTCTTCA GAGGACTCCACTTCA

CTCCTGAGGAGAAGT CTCCTGAGGTGAAGT
Probe for normal gene Probe for disease gene
 SNPs are abundant
• Estimated that 1 SNP occurs every 1000-3000
bp along the DNA of every chromosome
• Over 1.4 million SNPS identified to date on
human chromosome.
• When SNPs occur in a gene that codes for a
body function, a disease can result.
• Pharmaceutical companies are cataloguing the
chromosomal locations of SNPs
 stopped

Identifying sets of
disease genes by
microarrays
 Testing issues
• Should we test people for genetic conditions
for which no cure exists?
• What are the accepted consequences if a
parent learns their unborn child has a genetic
defect?
• What are the psychological consequences of
a false results that indicates that a healthy
person has a disease gene or a gene defect?
• How do we ensure privacy and
confidentiality?
Pharmacogenomics
Microarray for Leukemia screening
 Drug delivery
• Getting drug to target organs and tissue
– Oral drug to treat arthritis in knee is not very efficient
– Drug solubility may be an issue
• Microspheres

Insulin delivered as
a powder through
an inhaler
 10-9 meters

1 meter
Nanomedicine
• Nanometer is one
billionth of a meter
– May be used for
delivery of small
sensors to target sites
in body
– Unclogging arteries
– Detect and destroy
cancer cells
 Artificial blood
• Cell-free solutions containing molecules that can
bind and transport oxygen like hemoglobin
• Benefits
– Disease-free alternative to real blood
– Constant supply
– Universal donor type
• Disadvantages
– Cannot perform all the functions of a red blood cell-
only oxygen delivery
• Source of iron
• Carbon dioxide removal
 A B A,B O

You Can Give Blood You Can Receive

Type
To Blood From

A+ A+ AB+ A+ A- O+ O-

O+ O+ A+ B+ AB+ O+ O-
Out of 100 donors . . . . .
B+ B+ AB+ B+ B- O+ O-

AB+ AB+ Everyone 16 donors

84 donors
are RH-
A- A+ A- AB+ AB- A- O- are RH+
O- Everyone O-
38 are O+ 7 are O-
34 are A+ 6 are A-
B- B+ B- AB+ AB- B- O-
9 are B+ 2 are B-
AB- AB+ AB- AB- A- B- O-
3 are AB+ 1 is AB-
Monoclonal antibodies
 stopped Gene therapy
• Delivery of therapeutic genes into the body to
correct disease conditions created by faulty gene
• How is it done?

2 4

3
Severe combined immune deficiency

Mutation in gene encoding

adenine deaminase enzyme
Shows up as incompetent
T-cells which prevent B-cells
from making antibodies
 Challenges

• How long will introduced genes remain in

body?
• What vectors to use?
– Viruses (adenovirus, influenza virus, herpes, HIV
• Will depend on what tissues you want to target
 Cell and tissue transplantation
• Parkinson’s disease
– Loss of brain cells that produce dopamine which is
the chemical that nerve cells use to communicate with
each other
• Results in tremors, loss of balance, dexterity, etc.
• Tissue damage
– Adult neurons do not regenerate like fetal neurons
• Fetal neuron transplants into damaged tissue has
resulted in partial recovery of neurological damage
• Recent studies indicate that not many survive at
the site of damage due to inflammatory response
of host and those that do, end up differentiating
into olfactory bulb neurons instead of medium-
sized spiny neurons
Cell and tissue transplantation

Drill hole
in skull
 Xenotransplantation
• Transplanting organs from one species into
another
• May someday become an alternative to human-
to-human transplantation
– 1984 baboon heart transplanted into a 12-year-old
human girl
• Girl died after 3 weeks as a result of organ rejection
– Can be avoided by matching immune system of donor
and acceptor
• Major histocompatibility complex
– Human leucocyte antigen (HLA) present on all of our cells
Pigs genetically engineered to lack a sugar-producing gene
that causes human bodies to reject pig organs
 Stem cells
• Embryonic stem cells
• Infant and adult stem cells
– Present in small numbers in
• Bone marrow
• Peripheral blood
• Skin epithelium
• Umbilical cord blood
• Dental pulp of infant’s teeth
– May be obtained by reprogramming somatic
cells
• Introduction of retroviruses carrying
reprogramming genes into fibroblasts
Embryonic Stem Cells

Subject stem
cells to specific
conditions to
encourage
differentiation
into one of
many cell types
 Reprogramming

retrovirus
What stages of early embryonic development are
important for generating embryonic stem cells?

Embryonic stem cells, as their name suggests, are derived

from embryos. Specifically, embryonic stem cells are
derived from embryos that develop from eggs that have
been fertilized in vitro in an in vitro fertilization clinic and
then donated for research purposes with informed consent
of the donors.

They are not derived from eggs fertilized in a woman's body.

The embryos from which human embryonic stem cells are

derived are typically four or five days old and are a hollow
microscopic ball of cells called the blastocyst.
Sources of adult and infant stem cells

Peripheral blood stem cells

Current NIH-supported research
• Umbilical cord stem cells are able to
repopulate the bone marrow of a small
child, but only a small number of cells are
obtained from each umbilical cord.

• Now seeking methods to expand cells in

culture to generate larger numbers for use
in clinical applications.
 NIH Supported Research
• Stem cells found in dental pulp of “baby
teeth” have the potential to become cells
expressing molecular markers characteristic
of dentin, bone, fat, and nerve cells.

• Advance: These cells could possibly be

used to repair damaged teeth, regenerate
bone, treat nerve injury or disease.
 Using stem cells
Step 1- Define the problem.
Brain and spinal cord injuries result in damage to neurons.
Damaged neurons are not replaced by the right kind of newborn
neurons

Parkinson’s Disease is a genetic defect that results in dead nerve

cells that do not produce sufficient amounts of dopamine needed
for normal nerve signaling
 Possible solution: introduce stem cells to
affected area of the brain
Step 2: Determine source of stem cells
Stem cells from another normal individual or stem cells from patient?

Step 3: Determine source of stem cells

• Blastocyst embryonic stem cells.
– pluripotent stem cells-most universal type
• Fetal stem cells.
– pluripotent stem cells
• Umbilical cord blood stem cells.
– multipotent stem cells-can become many
different types of cells but natural fate is
to become blood and immune cells
• Adult stem cells.
– multipotent adult stem cells but best to
use ones that are destined for specific
functions (dopamine production in brain)
Step 4: If source is from another individual, match the stem cells
with the transplant recipient

If source is from patient’s previously collected and stored

stem cells, use these

If patient has Parkinson’ Disease, then introduce good copy

of gene in chromosome of stem cells
STEP 5: Introduce stem cells at site of injury or where
function needs to be restored
 STEP 5: Make the transplanted cells perform
the function

The implanted cells must survive and differentiate into the proper nerve cells
(medium –sized spiny neurons in case of injury or dopamine-producing cells in
case of Parkinson’s Disease patient.
Some patients reported a lessening in the severity of their symptoms.

Some patients who underwent the procedure experienced severe side

effects, including involuntary muscle twitching and jerking.
 Cloning
• Can refer to a gene, a cell, or an entire
organism
• Reproductive vs. therapeutic cloning
– Goal of reproductive cloning is to create a
baby
– Goal of therapeutic cloning is to provide stem
cells that are a genetic match to a patient wh
requires a transplant.
• Differences are diagrammed in Table 11.3
of text
 Regulations and ethics related to
stem cell
• Regs are in a state of flux in this country
• Ethical issues and regulations are
intertwined
• In Chapter 12, pages 314-315 a historical
perspective is provided.
• Your generation will likely play a significant
role not only in the development of the
technology but also the rules governing
the application of the technology
Learning Objectives
After completing this chapter you should be able to:
•Provide examples of model organisms and explain why they are important.
•Describe different karyotyping techniques that can detect chromosome abnormalities
and molecular techniques for genetic testing.
•Provide examples of why pharmacogenomics can change how many genetic disease
conditions may be treated in the future.
•Discuss how monoclonal antibodies may be used for treating disease.
•Understand the purpose of gene therapy, and compare and contrast different gene
therapy strategies and recognize limitation of gene therapy.
•Define regenerative medicine and provide examples of how cell and tissue
transplantation and organ engineering can be used.
•Understand what stem cells are and describe how they can be isolated. Provide
examples of possible therapies that may be developed from stem cells in the future.
•Compare and contrast therapeutic cloning and reproductive cloning.
•Briefly explain how molecular biology techniques and the Human Genome Project are
being used to create human disease gene maps.
 Summary
• What is meant by “model organisms” in the
context of detecting and diagnosing human
disease?
• What is karyotyping and what is its purpose?
• How will pharmacogenomics change how many
genetic disease conditions may be treated in the
future?
• How can monoclonal antibodies be used for
treating disease?
• What is purpose of gene therapy? What are its
different forms? What are its limitations?
• Describe how cell and tissue transplantation and
organ engineering can be used in regenerative
medicine
 Summary
• How are stem cells obtained? Provide
examples of possible therapies that may
be developed from stem cells in the future.
• What is the difference between therapeutic
cloning and reproductive cloning.
• How are molecular biology techniques and
the Human Genome Project being used to
create human disease gene maps?