FUNCTIONAL CURE OF HIV USING CRISPR
-ADLENE JENITTA, VIKRANT THOKCHOM, W. JASMINE
KARUNYA INSTITUTE OF TECHNOLOGY AND SCIENCES, COIMBATORE
INTRODUCTION
The human immunodeficiency virus (HIV) is a lentivirus (a
subgroup of retrovirus that causes HIV infection and over
time acquired immunodeficiency syndrome (AIDS).[1][2] AIDS is
a condition in humans in which progressive failure of
the immune system allows life-threatening opportunistic
infections and cancers to thrive. Without treatment, average
survival time after infection with HIV is estimated to be 9 to
11 years, depending on the HIV subtype. Human
immunodeficiency virus-1 (HIV-1) infection
continues to be a major health issue with more than
35 million individuals being infected worldwide.
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WHAT IS CRISPR TECHNOLOGY?
CRISPR is an abbreviation of Clustered
Regularly Interspaced Short Palindromic
Repeats.
It is a family of DNA sequences in bacteria.
These sequences play a key role in a prokaryotic
defence system, and forms the basis of a
technology known as CRISPR/Cas9 that
effectively and specifically changes genes within
organisms. Due to this nature, it can be used as
a functional cure for HIV.
Combination antiretroviral therapy (ART) has
been effective at suppressing viremia and
restoring primary host CD4+ T cells as well as
improving clinical outcomes, thereby
transforming a deadly infection into a
manageable chronic disease. However, low-level
virion production continues to exist due to viral
persistence inside cellular and anatomical
reservoirs. Over the last two decades, advances
have been achieved in genome editing
technology. However difficulties associated with
design, synthesis, and protein validation for a
specific gene locus of interest have restricted the
feasibility of these methods. A key breakthrough
was made when a bacterial immune system-
related RNA molecule, called clustered regularly
interspaced short palindromic repeats(CRISPR)
was found to be able to guide CRISPR-
associated 9 (Cas9) nuclease towards DNA
sequences matching those of the guide RNA
(gRNA)
The CRISPR/Cas9 system provides a novel and
promising tool for editing the HIV-1 proviral
genome.
CRISPR/Cas9 system targets the regulatory
genes of HIV-1 and inhibits viral replication in
infected T-cell cultures.
The experiment targeted two of HIV’s nine genes that
are key to the proliferation of the virus. The genes in
question are known as tat and rev. Without those
genes, HIV doesn’t work. Researchers used genetic
information from six common HIV subtypes to build
guide RNA (gRNA) that pointed the Cas9 protein at
the right DNA segments. They introduced it into cells
using a modified lentiviral vector.
They created a lentiviral vector that expresses Cas9
and gRNA. When they introduced this vector to
cultured cells that expressed the regulatory gene
products Tat and Rev, they succeeded in significantly
lowering the expression and functions of both Tat and
Rev. The team found no off-target mutations (non-
specific genome editing that unintentionally targets
the host cell genes), and the expression of Cas9 and
gRNA did not affect the survival rate of the cultured
cells.
By introducing gRNA and Cas9 to cultured cells with
a latent or persistent HIV-1 infection, they were able
to markedly suppress cytokine-dependent HIV-1
reactivation in latently infected cells and HIV-1
replication from persistently infected cells.
Furthermore, by introducing all six types of gRNA at
the same time, they managed to almost completely
block virus production from the infected cells.
OTHER ASPECTS
 Drugs of the new age:
CRISPR/Cas 9 could be a revolution for the
pharmaceutical industry. New drugs to treat
heart disease, blood disorders, and blindness
are recently announced which focuses more on
genetic methods.
 The arrival of Superplants:
Researchers are currently experimenting
with ways to improve crop disease resistance
and environmental stress tolerance using the
gene-editing tool.
 Boosting Human Intelligence:
CRISPR principle could be used to boost
the expected intelligence of an embryo by a
considerable amount.
 Designing babies:
One can sit and decide how the kid should
look like and what traits should he or she have.
 Eternal youth and living forever:
With this technology, diseases can be
eradicated and perfectly healthy humans can be
designed.
REFERENCES
tein (cyan
CRISPR-associated complex for
andefense) tiviral
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