CRISPR-The trend

of tomorrow !
 It is short for Clustered Regularly Interspaced
Short Palindromic Repeats (CRISPR). It is a
medium for genome editing in cells and
tissues.

It was first found in prokaryotic species as a

defense mechanism against the invading viruses.
Specifically, as the virus invades by transferring
a part of its DNA in the bacteria, CRISPR
mechanism comes into action! It first copies the
viral DNA and makes it's complimentary
sequence. Then this RNA sequence binds with a
Cas9 protein which can cut DNA, the RNA helps
the Cas9 protein to locate the viral DNA, which
then cuts thus disabling the virus!

This led the scientists to find something even

interesting. They found that the Cas9 protein not
only cuts the viral DNA but can also cut DNA
strands of other living beings!

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 How does it work
Basically, it is a segment of DNA comprising of
the short palindromic repeats and in between
those repeats lies what we call the spacer DNA,
now what makes the spacer DNA so unique is
that they are all different! More specifically they
are the history of the viral DNA that the system
uses to locate where the cut has to be made.

In the system, there are genes associated with

CRISPR called the Cas genes. What these genes
do is that they make the Cas proteins.

Now what happens is that the spacer DNA and

the one sequence of the repeat called the pre
crRNA (pronounced Precursor CRISPR RNA)
binds with the Cas9 protein and the tracrRNA
(trans-activating crRNA) which helps in locating
the gene in the DNA where the cut has to be
made and the Cas9 protein cuts it thus making
the DNA inactivate.

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 Now what scientists did is the bonded the
tracrRNA and the crRNA and made a so called
‘chimera’ they call it gRNA (guideRNA). Which
made it possible to alter the DNA.

But they didn’t stop here, they then introduced a

Host RNA which when the cell tries to fir the cut
in the DNA, binds with the DNA in the place of
the cut so now it was possible not only to alter it
but also change it!

It works!
It is possible to make large scale changes even in
complex living beings and not just in theory but
it has been tried and the experiment was
successful!

To test the strength of this mechanism, it was

tested on a couple of black mices to turn them

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 into albino by targeting tyrosinase (-an enzyme
which controls the production of melanin.

CRISPR is the key!

This mechanism is so accurate that it can literally
erase the disease! As this alters the body
genetically, it would be as if the disease was
never there. It can really help us in eradicating
the biggest rivals of all time.
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 Cancer:
We all know cancer is just an error or a mutation
in our genes which goes on multiplying and
spreading from the origin. Now there are some
genes which try to stop this uncontrollable
growth of cells by recognizing the mutations
trying to repair it (viz. Tumour suppressor genes,
DNA repair genes, etc.). But the main problem
arises when the error is in these genes. This can
be resolved by using CRISPR technology,
CRISPR can change the mutation and correct it
or even enhance by introducing the host RNA,
thus helping the immune system to fight better!

HIV:
Basically, the HIV virus attacks the T-
lymphocytes (aka CD4 cells) which play an
important role in our immune system. The virus

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 not only attacks and kills them but also by using
its replicating system turns the CD4 cells into the
virus! This reduces the count of these WBCs and
thus degrading our immune system. But CRISPR
can turn the whole thing upside down. Its Cas
Class I protein can copy the viral DNA which
can bind with the Cas9 protein which will
located and cut the viral DNA with the help of
gRNA and thus disable the HIV virus! And again
by introducing the host RNA we can enhance
and increase the count of CD4 cells.

Sickle Cell Anaemia:

This arises due to a particular mutation in a
specific gene called Haemoglobin Beta gene
which make the RBCs rigid, sticky and sickle
shaped, which then get stocked in the vessels and
block them and eventually making the patient
oxygen deficient.

The mutation in the gene is that instead of a

adenine-thymine pair it is thymine-adenine pair

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 and after transcribing the mRNA consists of a
uracil molecule instead of a adenine molecule.
And the protein is formed after translating,
instead of glutamic acid which is necessary
valine is formed.

This one mutation results in severe pain and in

most of the cases its fatal!

But CRISPR, can help this. It can alter the DNA

at the desired position and by the help of the host
RNA, it can change the thymine-adenine pair to
adenine-thymine pair. Thus resulting in the
formation of glutamic acid, and keeping the
RBCs healthy, flexible, smooth and circular.

It’s Healthy!
Due to population expansion, although
conventional breeding has increased crop yield it
is not so efficient as it is often accompanied by
loss of fitness and genetic diversity and moreover
it is rather time consuming, so it doesn’t ensure
sufficient food supply.
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 This makes the need for increasing crop yield
urgent, and the factors that limit this are biotic,
abiotic stresses, etc.

Genetic modification by CRISPR can resist these

stresses and increase the tolerance and quality of
the crop.

Resistance to biotic stress:

Biotic stresses include attack of viruses, bacteria,
pests, insects, etc. Out of these, given its minute
size, tackling virus is the most crucial. With the
help of CRISPR mechanism, two strategies can
be used for virus, one consists of designing a
sgRNA (single guide RNA) that targets viral
genome directly through sequence
complementation. Other consists of designing a
crop that confers antiviral characteristics!

Resistance to abiotic stress:

According to Charles Darwin’s theory of
evolution, we know it is not the most intellectual

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 but who can adapt to change is the one that
survives and keeps procreating. Due to climate
change, drought, floods, heat and cold genetic
modification ensures better yield.

Quality of the crops:

Net quality depends on both external as well as
internal quality.

External quality depends upon the colour, size

and the texture of the yield. It was found that, by
targeting a transferase enzyme named Phytoene
synthase (also used in the biosynthesis of
carotenoids which give characteristic colour to
the yield) and anthocyanin (a pigment which
changes its colour according to its pH).

Internal characteristics depend on the amount of

carbohydrates, minerals, vitamins, proteins, fats,
etc. that comprises the yield. They also depend
upon the level of nutrients and the amount of
bioactive compounds (they are defined as the
extra-nutritional compounds that occur in small
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 quantities in our food and can prevent or reduce
the risk of cardiovascular diseases and certain
types of cancer). Some bioactive compounds
such as lycopene, anthocyanin, resveratrol,
Gamma-aminobutyric acid, anthocyanin, indoles,
tannins, malate, etc. These are present in the
crops and their quantities can be increased by
CRISPR! Resulting in a better and a healthier
yield.

Thus, it can be established that CRISPR really

will be a trend in the near future!

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