Multiplexed orthogonal genome editing and transcriptional activation by Cas12a

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.The length of the guide RNA for Cas12a-VPR determines whether a target gene is edited or activated and allows for multiplexed, combinatorial gene modifications.

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