Synthetic Type III-E CRISPR-Cas Effectors for Programmable RNA-targeting.

Abstract

The recent discovery of the type III-E class of CRISPR-Cas effectors has reshaped our fundamental understanding of CRISPR-Cas evolution and classification. Type III-E effectors are composed of several Cas7-like domains and a single Cas11-like domain naturally fused together to create a single polypeptide capable of targeting and degrading RNA. Here we identified a novel type III-E-like effector composed of three Cas7 domains and a Cas1 domain which was not active but could be engineered into an active chimeric RNA-targeting Cas effector by domain additions and swaps from other type III-E effectors. The results reveal that various domains in type III-E effectors can be swapped for the equivalent domain from a different type III-E effector. Remarkably, the Cas1 domain located at the C-terminus of Cas7-1 could be swapped in place of the Cas11 domain located between the Cas7.1 and the Cas7.2 domains of DiCas7-11. The results reveal a new modality for engineering type III-E effectors from the blueprints found in nature.

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laboratory

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published

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Conflict statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Omar S. Akbari reports a relationship with Synvect Inc. that includes: equity or stocks. Omar S. Akbari reports a relationship with Agragene Inc. that includes: equity or stocks. Daniel J. Brogan has patent pending to University of California San Diego. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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