CRISPR-Cas9-mediated Adaptive Immunity Mechanism in Bacteria [3 Steps of CRISPR-Cas system]

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What is CRISPR-Cas9? How does CRISPR-Cas9 work in its natural context? How does CRISPR-Cas9 protect bacteria against pathogens by providing adaptive immunity? What are the different types of CRISPR systems? CRISPR-Cas9 is a Type II-A CRISPR system. In general, this video explains the mechanism of CRISPR-Cas9 function. Specifically, this video explains the three stages of CRISPR mediated pathogen defense: adaptation, crRNA biogenesis, and interference. The adaptation stage is used to acquire the spacer from the pathogen for insertion into the CRISPR locus. The crRNA biogenesis retrieves the spacer from the CRISPR locus in form a Cas nuclease. The Interference stage brings the Cas nuclease to the pathogen DNA for the RNA guided DNA cleavage resulting in a double-stranded DNA break upstream of the PAM region. There are many proteins/RNAs (cas1, cas2, csn2, and tracrRNA) and additional steps involved in this intricate 3-step pathway (like PAM scanning, PAM verification, protospacer trimming, maturation of crRNA, etc.). This video details some of the important steps in CRISPR-Cas mediated immunity and DNA cleavage.

00:00 Bacterial Immunity
01:14 CRISPR-Cas9 locus structure
02:42 Three steps CRISPR-Cas9
03:35 CRISPR-Cas Types
04:23 CRISPR Adaptation Spacer
08:07 crRNA biogenesis
10:51 Cas9 Interference
12:55 Cas9 and DNA cleavage

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All videos on Genetic Engineering: https://www.youtube.com/playlist?list=PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
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