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.