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CRISPR-Cas9 UPSC NOTE

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  Why in news A CRISPR system built to use the FnCas9 enzyme was found to edit genomes more efficiently and with less unintended damage th...

 Why in news

  • A CRISPR system built to use the FnCas9 enzyme was found to edit genomes more efficiently and with less unintended damage than existing technologies, researchers at CSIR-Institute of Genomics and Integrative Biology, New Delhi

CRISPR-Cas9

  • Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a gene editing technology, which replicates natural defence mechanism in bacteria to fight virus attacks, using a special protein called Cas9.

  • It usually involves the introduction of a new gene, or suppression of an existing gene, through a process described as genetic engineering.

  • CRISPR technology does not involve the introduction of any new gene from the outside.

  • CRISPR-Cas9 technology is often described as ‘Genetic Scissors’.

  • Its mechanism is often compared to the ‘cut-copy-paste’, or ‘find-replace’ functionalities in common computer programmes.

  • A bad stretch in the DNA sequence, which is the cause of disease or disorder, is located, cut, and removed and then replaced with a ‘correct’ sequence.

  • The tools used to achieve this are biochemical i.e., specific protein and RNA molecules.

  • The technology replicates a natural defence mechanism in some bacteria that uses a similar method to protect itself from virus attacks.

Mechanism:

  • The first task is to identify the particular sequence of genes that is the cause of the trouble.

  • Once that is done, an RNA molecule is programmed to locate this sequence on the DNA strand

  • After this Cas9 is used to break the DNA strand at specific points, and remove the bad sequence.

  • A DNA strand, when broken, has a natural tendency to re-attach and heal itself. But if the auto-repair mechanism is allowed to continue, the bad sequence can regrow.

  • So, scientists intervene during the auto-repair process by supplying the correct sequence of genetic codes, which attaches to the broken DNA strand.

  • The entire process is programmable, and has remarkable efficiency, though the chances of error are not entirely ruled out.

Limitations

  • CRISPR-Cas9 system can also recognise and cut parts of the genome other than the intended portion. 

  • Such “off-target” effects are more common when using the SpCas9 enzyme derived from Streptococcus pyogenes bacteria. 

  • Scientists have been able to engineer versions of SpCas9 with higher fidelity but only at the cost of editing efficiency

FnCas9 enzyme

  • To overcome these issues, researchers are exploring Cas9 enzymes from Francisella novicida bacteria. 

  • While this Cas9, called FnCas9, is highly precise, it has low efficiency as well.

  • Researchers tinkered with amino acids in FnCas9 that recognise and interact with the protospacer adjacent motif (PAM) sequence on the host genome. 

  • By doing this, increase the binding affinity of the Cas protein with the PAM sequence

  • The Cas9 can then sit on the DNA in a stronger configuration, and your gene editing becomes much more effective.

Potential applications

  • Enhanced FnCas9 is a viable option for treating genetic disorders

  • Enzyme’s efficient at correcting a genetic mutation that causes Leber congenital amaurosis type 2 (LCA2), a form of inherited blindness. 

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Learnerz IAS | Concept oriented UPSC Classes in Malayalam: CRISPR-Cas9 UPSC NOTE
CRISPR-Cas9 UPSC NOTE
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