The CRISPR-Cas9 system

is a tool for cutting DNA

at a specifically targeted location.
The technique has already revolutionized gene editing
but scientists are always looking
for new possibilities,
so what else can CRISPR do?

Since being discovered in a bacterial immune system

CRISPR-Cas9 has been adapted
into a powerful tool for genomic research.

There are two components to the system:

a DNA-cutting protein called Cas9
and an RNA molecule known as the guide RNA.

Bound together, they form a complex

that can identify and cut specific sections of DNA.

First, Cas9 has to locate and bind

to a common sequence in the genome called a PAM.

Once the PAM is bound,

the guide RNA unwinds part of the double helix.

The RNA strand is designed

to match and bind a particular sequence in the DNA.
Once it’s found the correct sequence,
Cas9 can cut the DNA –
its two nuclease domains each make a nick
leading to a double strand break.
Although the cell will try to repair this break,
the fixing process is error-prone
and often inadvertently introduces mutations
that disable the gene.
This makes CRISPR a great tool
for knocking out specific genes.

Several labs have been working on ways to use CRISPR

to promote gene transcription.
They do this by deactivating Cas9 completely
so it can no longer cut DNA.
The same principle applies to gene silencing.
A KRAB domain fused to the Cas9
inactivates transcription by recruiting more factors
that physically block the gene.
A more outside-the-box idea for using CRISPR
is to attach fluorescent proteins to the complex
so you can see where particular DNA sequences
are found in the cell.
This could be useful for things like visualizing
the 3D architecture of the genome,
or to paint an entire chromosome
and follow its position in the nucleus.
CRISPR has already changed the face of research
but these new ideas show
that what’s been achieved so far
could just be the tip of the iceberg
when it comes to CRISPR’s potential.
Whatever comes next,
it seems the CRISPR revolution is far from over.