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Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife

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Grace A. Meaker,Emma J. Hair,

Thomas E. Gorochowski^*

Grace A. Meaker,Emma J. Hair,

Thomas E. Gorochowski^*

This version is not peer-reviewed

Submitted:

30 September 2020

Posted:

01 October 2020

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Abstract

The RNA-guided endonuclease system CRISPR-Cas9 has been extensively modified since its discovery, allowing its capabilities to be extending far beyond double-stranded cleavage to high fidelity insertions, deletions, and single base edits. Such innovations have been possible due to the modular architecture of CRISPR-Cas9 and the robustness of its component parts to modifications and the fusion of new functional elements. Here, we review the broad toolkit of CRISPR-Cas9-based systems now available for diverse genome editing tasks. We provide an overview of their core molecular structure and mechanism and distil the design principles used to engineer their diverse functionalities. We end by looking beyond the biochemistry and towards the societal and ethical challenges that these CRISPR-Cas9 systems face if their transformative capabilities are to be deployed in a safe and acceptable manner.

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Subject: Biology and Life Sciences - Biochemistry and Molecular Biology

Copyright: This open access article is published under a Creative Commons CC BY 4.0 license, which permit the free download, distribution, and reuse, provided that the author and preprint are cited in any reuse.