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. 2019 Jan;16(1):51-54.
doi: 10.1038/s41592-018-0262-1. Epub 2018 Dec 17.

Multiplexed orthogonal genome editing and transcriptional activation by Cas12a

Marco Breinig  1 , Anabel Y Schweitzer  1 , Anna M Herianto  1 , Steffie Revia  1 , Lisa Schaefer  1 , Lena Wendler  1 , Ana Cobos Galvez  1 , Darjus F Tschaharganeh  2
Affiliations

Multiplexed orthogonal genome editing and transcriptional activation by Cas12a

Marco Breinig et al. Nat Methods. 2019 Jan.

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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