Make was Cas9, there's a new pair of genomic
scissors in town and its name is Cpf1!
If you're a CRISPR fan, you're probably familiar
with the Cas9 nuclease, the revolutionary
tool that is totally democratizing genetic
engineering as we know it. Since the discovery
of its applications, scientists have been
racing to develop new and improved CRISPR
tools to take genomic editing to the next
level.
One of the latest CRISPR innovations that
is getting the scientific world all abuzz
with excitement is the discovery of Cpf1,
a new CRISPR nuclease that is coming soon
to Applied Biological Materials.
This is Molecular Minutes, bringing you breakthroughs
at breakneck speeds and in this video we'll
take you to the cutting edge of CRISPR technologies
and give you the lowdown on what soon may
be a rising star in genomic editing.
First characterized in 2015 by the Zhang lab
at MIT, Cpf1 was picked out of hundreds of
potential CRISPR systems used by a wide range
of bacterial species. Sleeker, simpler and
even more versatile than the current star
of CRISPR, Cpf1 promises to enable a whole
new level of precision genomic editing, empowering
scientists to break new ground in their research.
Here's why:
Cpf1 doesn't require tracrRNA, it only requires
CRISPR RNA, reducing the size of the engineered
gRNA molecule required by half compared to
Cas9, making genome editing cheaper
than ever before! That means a whole lot more
money for beer!
Cpf1 itself is also smaller than Cas9! And
size matters when it comes to gene delivery.
Combined with its shorter gRNA, Cpf1 is even
easier to shuttle into cells via low capacity
vectors such as the adeno-associated virus.
While Cas9 generates blunt ends after cutting,
Cpf1 clips DNA in such a way that it leaves
behind sticky 5' overhangs. Researchers can
design DNA inserts that dock perfectly on
these overhangs, optimizing the non-homologous
end joining repair pathway for DNA insertion.
This means that with only one pair of scissors,
Cpf1 can get even a cut above Cas9 in non-dividing
cell types that rely mostly on this NHEJ repair
pathway.
Cpf1 also out-snips Cas9 because it cuts a
comfortable distance away from its PAM site.
This means that cutting does not disrupt the
PAM site, allowing for multiple rounds of
DNA cleavage that promise to increase opportunities
for the desired genomic editing to occur.
This is in comparison to Cas9 which cuts closer
to its PAM site resulting in InDels that destroy
the recognition sequence and prevent further
rounds of cutting.
Finally, for those looking to explore and
edit new realms of the genome not covered
by Cas9, Cpf1 might just be your ticket! Whereas
Cas9 relies on G-rich PAM sequences, Cpf1
recognizes T-rich PAM sequences, significantly
expanding the breadth of possible genomic
targets.
Zhang and his team have already identified
two Cpf1 orthologs that can carry out effective
genome editing in human cells, opening up
new avenues for research and therapeutic applications.
With the Cas9 and Cpf1 CRISPR dream team at
our disposal, the future of genomic editing
seems limitless! Are you as excited as we
are?
We can't wait to let you try this new CRISPR
tool so we're working hard to make it available
at abm as soon as possible. In the meantime,
you can still get up close and personal with
your favorite genes using our suite of CRISPR
Cas9 nucleases, nickases, and null mutants.
These are available as AAV or lentiviral vectors
or viruses. We even do custom cell line knockouts
in whatever cell line you want. Now that's
genome editing made easy.
Thanks for watching Molecular Minutes, stay
tuned for more newsflashes as we follow the
latest movers and shakers in the biotech industry.
