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

Grace A. Meaker; Emma J. Hair; Thomas E. Gorochowski

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preprints.org > biology and life sciences > biochemistry and molecular biology > 202007.0351.v2

Working Paper Review Version 2 This version is not peer-reviewed

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

Grace A. Meaker

Emma J. Hair

Thomas E. Gorochowski

Version 1 : Received: 15 July 2020 / Approved: 16 July 2020 / Online: 16 July 2020 (12:59:04 CEST)
Version 2 : Received: 30 September 2020 / Approved: 1 October 2020 / Online: 1 October 2020 (08:38:22 CEST)

How to cite: Meaker, G.A.; Hair, E.J.; Gorochowski, T.E. Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife. Preprints.org 2020, 2020070351 Meaker, G.A.; Hair, E.J.; Gorochowski, T.E. Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife. Preprints.org 2020, 2020070351

Cite as:

Meaker, G.A.; Hair, E.J.; Gorochowski, T.E. Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife. Preprints.org 2020, 2020070351 Meaker, G.A.; Hair, E.J.; Gorochowski, T.E. Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife. Preprints.org 2020, 2020070351

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.

Keywords

synthetic biology; CRISPR; Cas9; biotechnology; biodesign; nickase; base editing; prime editing; genome editing; ethics; responsible innovation

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Comments (1)

Comment 1

Received: 1 October 2020
Commenter: Thomas Gorochowski
Commenter's Conflict of Interests: Author

Comment: We have expanded on our description of several of the systems and included a new figure covering the basic function of the CRISPR-Cas9 system and table showing its breadth of use.

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