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TitleStructural basis for self-cleavage prevention by tag:anti-tag pairing complementarity in type VI Cas13 CRISPR systems.
Journal, issue, pagesMol Cell, Vol. 81, Issue 5, Page 1100-11115.e5, Year 2021
Publish dateMar 4, 2021
AuthorsBeibei Wang / Tianlong Zhang / Jun Yin / You Yu / Wenhao Xu / Jianping Ding / Dinshaw J Patel / Hui Yang /
PubMed AbstractBacteria and archaea apply CRISPR-Cas surveillance complexes to defend against foreign invaders. These invading genetic elements are captured and integrated into the CRISPR array as spacer elements, ...Bacteria and archaea apply CRISPR-Cas surveillance complexes to defend against foreign invaders. These invading genetic elements are captured and integrated into the CRISPR array as spacer elements, guiding sequence-specific DNA/RNA targeting and cleavage. Recently, in vivo studies have shown that target RNAs with extended complementarity with repeat sequences flanking the target element (tag:anti-tag pairing) can dramatically reduce RNA cleavage by the type VI-A Cas13a system. Here, we report the cryo-EM structure of Leptotrichia shahii LshCas13a in complex with target RNA harboring tag:anti-tag pairing complementarity, with the observed conformational changes providing a molecular explanation for inactivation of the composite HEPN domain cleavage activity. These structural insights, together with in vitro biochemical and in vivo cell-based assays on key mutants, define the molecular principles underlying Cas13a's capacity to target and discriminate between self and non-self RNA targets. Our studies illuminate approaches to regulate Cas13a's cleavage activity, thereby influencing Cas13a-mediated biotechnological applications.
External linksMol Cell / PubMed:33472057 / PubMed Central
MethodsEM (single particle)
Resolution3.06 Å
Structure data

30767

7dmq

EMDB-30767, PDB-7dmq:
Cryo-EM structure of LshCas13a-crRNA-anti-tag RNA complex
Method: EM (single particle) / Resolution: 3.06 Å

Source
  • leptotrichia shahii (bacteria)
  • escherichia coli (E. coli)
KeywordsIMMUNE SYSTEM/RNA / Type VI-A CRISPR-Cas system / Cas13a / anti-tag RNA / inhibition / IMMUNE SYSTEM / IMMUNE SYSTEM-RNA complex

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