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Advances in Genetic Technology Class Notes Make sure you study this along with our first PowerPoint on Transgenics and your class Article notes

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Title: Advances in Genetic Technology Class Notes Make sure you study this along with our first PowerPoint on Transgenics and your class Article notes


1
Advances in Genetic Technology Class Notes Make
sure you study this along with our first
PowerPoint on Transgenics and your class Article
notes
2
When we talk about cloning, are we always talking
about the same thing?
3
  • recombinant DNA cloning (transfer of gene of
    interest to a vector like bacteria or a virus
    which will incorporate in all future DNA)
  • reproductive cloning (transfer of nuclear
    material from a somatic cell of one organism to
    an egg of another that has had its nucleus
    removed, resulting in an offspring like the
    donor)
  • therapeutic cloning (also known as embryo cloning
    is designed to create stem cells for use in
    treating disease)

4
  • How is a mammal cloned (reproductive cloning)?

5
Is this possible In humans?
There are lots of efficiency problems. It takes
hundreds of tries to get one clone, even in a
sheep.
6
What is PCR and why do we need it?
  • Polymerase chain reaction (PCR) is a fast and
    inexpensive technique used to "amplify" - copy -
    small segments of DNA.
  • It is used wherever large amounts of DNA are
    needed. Most often there is only a small sample
    of DNA to begin with.
  • Examples of its use DNA fingerprinting,
    detection of bacteria or viruses (particularly
    AIDS), and diagnosis of genetic disorders.

7
Steps to PCR mimic what happens in replication
8
If you run about 35 cycles and end up with
billions of copies of the original DNA
9
  • What is a Short Tandem Repeat (STR)Polymorphism
    and how is it used?
  • STRs are short sequences of DNA, normally of
    length 2-5 base pairs, that are repeated numerous
    times.
  • These are inherited regions of our DNA that can
    vary from person to person.
  • These are located at the same place on the
    chromosomes but they vary (polymorphism) in
    length from person to person.
  • It is highly unlikely that an individual would
    have the same number of repeats as another
    individual at the most common13 STR sites used by
    criminal justice to distinguish individuals.

10
These loci on the chromosomes are the location of
STRs and are used to distinguish one individual
from another. PCR can be used to make many copies
of the DNA segments with STRs for forensic
analysis.
11
Electrophoresis is used in separating DNA
fragments
  • Gel electrophoresis is a method of separating
    large molecules (such as DNA fragments or
    proteins) from a mixture of similar molecules.
  • An electric current is passed through a gel
    containing the DNA mixture, and each kind of
    molecule travels through the medium at a
    different rate, depending on its electrical
    charge and size.
  • The gel acts as a sieve. DNA moves at different
    rates because larger molecules move through the
    gel matrix slower than smaller molecules.

12
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13
The Human Genome Project (approx. 1990-2003)
  • Project goals were to
  • identify all the approximately 20,000-25,000
    genes in human DNA,
  • determine the sequences of the 3 billion chemical
    base pairs that make up human DNA,
  • store this information in databases,
  • improve tools for data analysis,
  • transfer related technologies to the private
    sector

14
Applications of the Data from the Human Genome
Project
  • Molecular medicine
  • Energy sources and environmental applications
  • Risk assessment
  • Bioarchaeology, anthropology, evolution, and
    human migration
  • DNA forensics (identification)
  • Agriculture, livestock breeding, and
    bioprocessing

15
What is Gene Therapy?
  • Gene therapy is designed to introduce beneficial
    versions of a gene into an afflicted individual
    for therapeutic purposes.
  • How does it work?
  • For gene therapy to be permanent, the cells that
    receive the normal allele must be ones that
    multiply throughout the patient's life.
  • In most gene therapy studies, a "normal" gene is
    inserted into the genome to replace an
    "abnormal," disease-causing gene using a vector.
  • Currently, the most common vector is a virus that
    has been genetically altered to carry normal
    human DNA.

16
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17
Two mechanisms direct into the tissue or
through the use of stem cells (unspecialized
cells) to carry the gene into the body.
18
How do small RNAs (microRNA, RNAi) impact the
expression of traits?
  • These are short nucleotide sequences that bind to
    the complementary RNA made during transcription
    and they usually silence the gene.
  • MicroRNAs target about 60 of all genes. They
    are abundant in all human cells and are able to
    repress hundreds of targets each.

19
When the microRNA interferes with translation
the protein will not be produced and therefore
the gene is silenced or not expressed.
20
How can we determine if a gene will express
itself?
With microarray technology.
A microarray is a tool for analyzing gene
expression that consists of a small membrane or
glass slide containing samples of many genes
arranged in a regular pattern.
Scientists can determine, in a single experiment,
the expression levels of hundreds or thousands of
genes within a cell by measuring the amount of
mRNA bound to each site on the array. Often
they are looking for Single Nucleotide
Polymorphisms (SNPs)
21
Compare a normal healthy sample with the patients
sample
  GREEN represents Control DNA,   RED represents
Sample DNA, where either DNA or cDNA is derived
from diseased tissue hybridized to the target
DNA.   YELLOW represents a combination of
Control and Sample DNA, where both hybridized
equally to the target DNA.   BLACK represents
areas where neither the Control nor Sample DNA
hybridized to the target DNA.  
22
Different types of microarrays have different
applications
In Brief Microarray Applications In Brief Microarray Applications
Microarray type Application
CGH Tumor classification, risk assessment, and prognosis prediction
Expression analysis Drug development, drug response, and therapy development
Mutation/Polymorphism analysis Drug development, therapy development, and tracking disease progression
23
What is a SNP?
  • It is a specific type of mutations in DNA. This
    small variation is enough to cause a disease or
    disability.
  • When researchers use microarrays to detect
    mutations or polymorphisms in a gene sequence,
    the target, or immobilized DNA, is usually that
    of a single gene.
  • In this case though, the target sequence placed
    on any given spot within the array will differ
    from that of other spots in the same microarray,
    sometimes by only one or a few specific
    nucleotides.
  • One type of sequence commonly used in this type
    of analysis is called a Single Nucleotide
    Polymorphism, or SNP, a small genetic change or
    variation that can occur within a person's DNA
    sequence.

24
Last but not least, what does epigenetics have to
do with this?
  • Epigenetics influence the expression of traits.
  • It is now clear that your destiny is not limited
    by your genes alone. There are plenty of
    non-genetic influences on expression of your
    traits.
  • It is through epigenetic marks that environmental
    factors like diet, stress and prenatal nutrition
    can make an imprint on genes that are passed from
    one generation to the next

25
  • Epigenetic mechanisms include methylation and
    acetylation of nucleotides which cause the DNA to
    bind tightly or more loosely around the histones.
  • The ultimate result is the transcription of
    information onto mRNA or not.

26
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