Crizelda D.

Liwanag, MSc, RMT

Genetic Manipulations

Applications in Diagnosis
 Applications in Diagnosis
1. Detection of DNA
• Electrophoresis
• Southern Blotting
• PCR
• DNA Fingerprinting
• DNA Sequencing
•3.
2. Chromosomes,
Each
The short piece
fragments which
in is used
a set range
asseparated
are a
in template
size from to50
byof generate
gel million toa 250
set of
electrophoresismillion
4. New
bases, fluorescent
fragments must
that
first dyes
differ allow
be broken
in lengthseparation
into
from
much
each all
shorter four fragments
other pieces
by a single in athat
(subcloning
base
(separation
single step).
be lane
step).
will on the
identified ingel.
a later step (template preparation and
sequencing reaction steps).

5.7. Automated
6. The
After
final
thebase
bases
sequencers
at the
are end
"read,"
analyze
of
each
the
computers
resulting
fragment are
electropherograms,
isused
identified
to
(base-calling
and
assemble
the outputthe
step).
isshort
a four-color
This
sequences
process
recreates
chromatogram
(in blocksthe of original
about
showing 500
sequence
peaks
basesthat
ofeach,
represent
As, Ts,called
each
Cs, the
andof read
the
Gs four
for
length)
each
DNA
short
bases.
into piece
long continuous
generated stretches
in the first
step.
that are analyzed for errors,
gene-coding regions, and other
characteristics.
Genetic Manipulations

Applications in Therapy
 Applications in Therapy
1. Mutagenecity and anti-
mutagenecity assays
2. Recombinant DNA technology
3. Gene transfer and movement
4. Gene therapy
5. RNA interference
6. Stem cell and organ cloning
7. GMO’s
Gene therapy
 Gene therapy

• Insert a working gene

 Gene therapy

• Use virus
 Gene therapy

• Like an envelope carrying a letter

 Gene therapy

• Gene randomly inserts itself into

genome
 Gene therapy

• It can now be read (correct

instruction)
RNA interference
 RNA interference

• Suppressors
 RNA interference

• From viruses (replace suppressors)

 RNA interference

• New genes cause cancer

 RNA interference

• Small interfering RNA binds to mRNA of

infected genes
 Stem cell therapy

• Stem cells
 Stem cell therapy

• Trick stem cells of a different organ into

regenerating an organ
 Stem cell therapy

• Damaged areas of the heart

 Stem cell therapy

• Black dots representing areas injected with

stem cells
 Stem cell therapy

• Stem cells from femur BM are transferred

to the heart and they mimic the cells
 Stem cell therapy

• Purple area shows healthy heart

 Stem cell therapy

• Frog’s eyes grown from stem cells

 Stem cell therapy

• 1st to grow sensory organs from embryonic

stem cells
 Stem cell therapy

• Frogs that cannot see are lighter in

color
 Stem cell therapy

• Clone of own embryo (to solve

histocompatibility and immunologic
tolerance problems)
Stem cell therapy
Embryonic stem cell cloning

• Co-creator of DOLLY
Embryonic stem cell cloning

• Trying to clone a human being

Believes cloning will be done
 What we still DON’T know, even with
the full human sequence in hand:
• Gene number, exact locations, and
functions
• Gene regulation
• DNA sequence organization
• Chromosomal structure and organization
• Noncoding DNA types, amount,
distribution, information content, and
functions
• Coordination of gene expression, protein
synthesis, and post-translational events
• Interaction of proteins in complex
molecular machines
 What we still DON’T know, even with
the full human sequence in hand:
• Predicted vs experimentally determined gene
function
• Evolutionary conservation among organisms
• Protein conservation (structure and function)
• Proteomes (total protein content and function) in
organisms
• Correlation of SNPs (single-base DNA variations
among individuals) with health and disease
• Disease-susceptibility prediction based on gene
sequence variation
• Genes involved in complex traits and multigene
diseases
• Complex systems biology including microbial
consortia useful for environmental restoration
• Developmental genetics, genomics