Structure-guided discovery of ancestral CRISPR-Cas13 ribonucleases-Reference-Cited by-同舟云学术

Structure-guided discovery of ancestral CRISPR-Cas13 ribonucleases

Published:2024-08-02 Issue:6708 Volume:385 Page:538-543
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Yoon Peter H.¹²³^ORCID,Zhang Zeyuan²³⁴⁵^ORCID,Loi Kenneth J.¹²^ORCID,Adler Benjamin A.²³⁵^ORCID,Lahiri Arushi¹²^ORCID,Vohra Kamakshi²⁵,Shi Honglue²³^ORCID,Rabelo Daniel Bellieny²⁵^ORCID,Trinidad Marena²³,Boger Ron S.²³⁴,Al-Shimary Muntathar J.¹²³^ORCID,Doudna Jennifer A.¹²³⁵⁶^ORCID

Affiliation:

1. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.

2. Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.

3. Howard Hughes Medical Institute, University of California, Berkeley, Berkeley CA, USA.

4. Biophysics Graduate Group, University of California, Berkeley, Berkeley, CA, USA.

5. California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA.

6. Gladstone Institutes, San Francisco, CA, USA.

Abstract

The RNA-guided ribonuclease CRISPR-Cas13 enables adaptive immunity in bacteria and programmable RNA manipulation in heterologous systems. Cas13s share limited sequence similarity, hindering discovery of related or ancestral systems. To address this, we developed an automated structural-search pipeline to identify an ancestral clade of Cas13 (Cas13an) and further trace Cas13 origins to defense-associated ribonucleases. Despite being one-third the size of other Cas13s, Cas13an mediates robust programmable RNA depletion and defense against diverse bacteriophages. However, unlike its larger counterparts, Cas13an uses a single active site for both CRISPR RNA processing and RNA-guided cleavage, revealing that the ancestral nuclease domain has two modes of activity. Discovery of Cas13an deepens our understanding of CRISPR-Cas evolution and expands opportunities for precision RNA editing, showcasing the promise of structure-guided genome mining.

Publisher

American Association for the Advancement of Science (AAAS)

Link

https://www.science.org/doi/pdf/10.1126/science.adq0553

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