Multiplexed orthogonal genome editing and transcriptional activation by Cas12a.

Breinig, Marco; Schweitzer, Anabel Y; Herianto, Anna M; Revia, Steffie; Schaefer, Lisa; Wendler, Lena; Cobos Galvez, Ana; Tschaharganeh, Darjus F · Nat Methods · 2019

basic_science · Level V

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Abstract

CRISPR-Cas9-based combinatorial perturbation approaches for orthogonal knockout and gene activation have been impeded by complex vector designs and co-delivery of multiple constructs. Here, we demonstrate that catalytically active CRISPR-Cas12a fused to a transcriptional-activator domain enables flexible switching between genome editing and transcriptional activation by altering guide length. By leveraging Cas12a-mediated CRISPR-RNA array processing, we illustrate that Cas12a-VPR enables simplified multiplexed knockout and transcriptional activation in vitro and in vivo.

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