A New Molecular biology technique for genome editing: CRISPR-Cas9 HOW NEW DISCOVERY IS CHANGING EVERYTHING AS WE KNOW IT. Vanessa Chappell Cell Seminar 2016
A New Molecular biology technique for genome editing: CRISPR-Cas9
HOW NEW DISCOVERY IS
CHANGING EVERYTHING AS
WE KNOW IT.
Vanessa ChappellCell Seminar 2016
Overview
• What is genome engineering and gene therapy?
• What molecular biology tools are available?
• What makes CRISPR a better option?
• Current research
• What does this mean for the future?
The basics:
. Gene Therapy
Experimental techniques used to fix a
genetic problem at its source instead
of using a drug or surgery
Genome editing
DNA is inserted, deleted or replaced
in the genome of an organism using
engineered nucleases
Programmable Nucleases
DNA targeting platforms for genome
editing and gene therapy
Programmable Nucleases
Zinc-finger nucleases (ZFNs)
● 1st genomic editing strategy
● recognizes 3-4 bases of
sequence
● cost is high
● “finiky” to make
● need a new finger for each
study
Transcription Activator-Like nucleases (TALENs)
● target a single nucleotide
● much larger than ZFN
● difficult to deliver
● fast to make
● cost is high (but less than ZFN)
Clusters Regularly Interspaced Short Palidromic Repeats (CRISPR)/CRISPR associated (Cas9)
● uses a short guide RNA
● this complex is NOT manmade
● easily modified
● inexpensive
● efficient
What makes CRISPR/Cas so special?
What is it?
● bacterial adaptive
immunity
● can be easily modified
and programmed to
target any organism's
DNA
Where did it come from?
It was discovered in bacteria
when a researcher noticed
that the spaces in between
the CRISPRs matched the
DNA of virus’ that targeted
the bacteria
What does it do?
upon attack or initiation it
uses stored bits of RNA to
target and destroy an
invader
UAB stem cell research lab 2016
The research team took stem cells from
a patient at Children’s of Alabama. They
converted these cells and used CRISPR-
Cas9 to target and correct the mutated
base pair. They were able to show that
the genes functioned normally using a
mouse model.
UAB researchers may be on their way to
curing sickle cell disease using CRISPR-
Cas9 technology.
***This research team is applying for
FDA permission to use what they have
found in the clinic on human patients.
Personalized MedicineBabies born in Alabama hospitals currently are
tested for 35 diseases, “but for about the same price,
we could sequence their genome” and pinpoint
genes that put them at risk for disease, Townes says.
Then, “long before someone develops one of these
genetic predispositions, we could correct the gene
so that they never experience that disease,” Townes
says.
Summary
In general CRISPR/Cas9 is better than other engineered nucleases
because:
• generally more efficient at editing than ZFNs and TALENs
• CRISPR is RNA based so no need to reengineer the proteins to
recognize new DNA being studied (like the others)
• it can be multiplexed by co-transfection or co-injection of gRNA
FUTURE RESEARCH
Medical
● treat genetic disease
● create specific antibiotics
● treat viral infections (HIV)
Agriculture
● GM plants
● pest resistant crops
● disease resistance livestock
Research
● study gene function
● target gene mutation
● create transgenic organisms
● synthetic biology
Why is it important?
Disease cure
● cystic fibrosis
● Huntington’s disease
● sickle-cell anemia
● HIV
● leukemia and other
blood disorders
● cancer
Agricultural
● better crops
● healthier livestock
● better breeding with
“gene drive”
Research
● faster methods
● less expensive
● more reliability
SETTING LIMITS ON WHAT SHOULD BE ALLOWED
• Barcode babies• Smart genes• Skinny genes• Pretty genes• What’s next??
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“
”“The first promise of any good politician is to make people's lives better and scientific research leading to Innovation is one of the best ways to honor that promise. Until about 1700, there was basically no development. Almost everybody was poor. Many were sick. One of every 4 children died. The average lifespan was about 40 years and 99% of people were illiterate. But then science came along and we started inventing---electricity, steam engine, antibiotics, sanitation, vaccines, microprocessors and genetic medicine…”
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