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Discovery and Characterization of Novel Type V Cas12f Nucleases with Diverse Protospacer Adjacent Motif Preferences by Allison Sharrar; Luisa Arake de Tacca; Trevor Collingwood; Zuriah Meacham; David Rabuka; Johanna Staples-Ager; Michael Schelle is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Discovery and Characterization of Novel Type V Cas12f Nucleases with Diverse Protospacer Adjacent Motif Preferences about?

Small Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR-associated (Cas) effectors are key to developing gene editing therapies due to the packaging constraints of viral vectors. While Cas9 and Cas12a CRISPR-Cas effectors have advanced into select clinical applications, their size is prohibitive for efficient delivery of both nuclease and guide RNA in a single viral vector. Type V Cas12f effectors present a solution given their small size. In this study, we describe a novel set of miniature (<490AA) Cas12f nucleases that cleave doublestranded DNA in human cells. We determined their optimal trans-activating RNA empirically through rational modifications, which resulted in an optimal single guide RNA. We show that these nucleases have broad protospacer adjacent motif (PAM) preferences, allowing for expanded genome targeting. The unique characteristics of these novel nucleases add to the diversity of the miniature CRISPR-Cas toolbox while the expanded PAM allows for the editing of genomic locations that could not be accessed with existing Cas12f nucleases.

Who reads Discovery and Characterization of Novel Type V Cas12f Nucleases with Diverse Protospacer Adjacent Motif Preferences?

Author

Allison Sharrar; Luisa Arake de Tacca; Trevor Collingwood; Zuriah Meacham; David Rabuka; Johanna Staples-Ager; Michael Schelle

Publisher

Mary Ann Liebert Inc

Published

2023

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)