Abstract
Microbes use diverse defence strategies that allow them to withstand exposure to a variety of genome invaders such as bacteriophages and plasmids. One such defence strategy is the use of RNA guided endonuclease called CRISPR-associated (Cas) 9 protein. The Cas9 protein, derived from type II CRISPR/Cas system, has been adapted as a versatile tool for genome targeting and engineering due to its simplicity and high efficiency over the earlier tools such as ZFNs and TALENs. With recent advancements, CRISPR/Cas9 technology has emerged as a revolutionary tool for modulating the genome in living cells and inspires innovative translational applications in different fields. In this paper we review the developments and its potential uses in the CRISPR/Cas9 technology as well as recent advancements in genome engineering using CRISPR/Cas9.
Highlights
Microbes use diverse defence strategies that allow them to withstand exposure to a variety of genome invaders such as bacteriophages and plasmids
In this paper we review the developments and its potential uses in the CRISPR/Cas9 technology as well as recent advancements in genome engineering using CRISPR/Cas9
The CRISPR region contains short repetitive base sequences separated by stretches of variable sequences referred to as spacers that share the sequence homology with foreign elements including bacteriophage and plasmid [4]
Summary
Bilal Ahmad Lone, Shibendra Kumar Lal Karna, Faiz Ahmad, Nerina Shahi, and Yuba Raj Pokharel. Microbes use diverse defence strategies that allow them to withstand exposure to a variety of genome invaders such as bacteriophages and plasmids One such defence strategy is the use of RNA guided endonuclease called CRISPR-associated (Cas) 9 protein. CRISPR immunity in microbes is acquired through (A) adaptation or spacer acquisition, (B) CRISPR-RNA (crRNA) biogenesis, and (C) target interference [5, 6] (Figure 1). During adaptation phase, invading DNA is spliced into small fragments and incorporated into a CRISPR locus as new spacers that become the memory record of infection. The PAM sequence is essential for selection and acquisition of protospaces into the CRISPR array by Cas and Cas protein complexes [10, 11] (Figure 1(A)).
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