Preprint Review Version 1 Preserved in Portico This version is not peer-reviewed

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

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 2020, 2020070351 (doi: 10.20944/preprints202007.0351.v1). Meaker, G.A.; Hair, E.J.; Gorochowski, T.E. Advances in Engineering CRISPR-Cas9 as a Molecular Swiss Army Knife. Preprints 2020, 2020070351 (doi: 10.20944/preprints202007.0351.v1).

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.

Subject Areas

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

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