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TitleDNA-guided CRISPR-Cas12a effectors for programmable RNA recognition and cleavage.
Journal, issue, pagesNat Biotechnol, Year 2026
Publish dateMay 1, 2026
AuthorsXiaolong Wu / Wai Hei Lam / Zibin Zhao / Yumeng Cao / Haosi Lin / Xianzhen Feng / Yuanliang Zhai / I-Ming Hsing /
PubMed AbstractCRISPR-Cas effectors typically rely on RNA guides to recognize target sequences. In Cas12a, the protospacer adjacent motif on DNA engages conserved protein residues, triggering target binding and ...CRISPR-Cas effectors typically rely on RNA guides to recognize target sequences. In Cas12a, the protospacer adjacent motif on DNA engages conserved protein residues, triggering target binding and nuclease activation. Here we reprogram Cas12a into a DNA-guided, RNA-targeting effector. Exploiting protospacer-adjacent motif-dependent interaction, we engineer synthetic CRISPR DNA that engages Cas12a to form a functional deoxyribonucleoprotein complex, while repurposing solely RNA as the programmable target. Structural, biophysical and biochemical analyses reveal the molecular basis of this DNA-guided, RNA-targeting configuration and support an activation pathway distinct from that of canonical RNA-guided systems. DNA-guided Cas12a enables direct RNA detection and efficient intracellular RNA knockdown, establishing a modular activation architecture for CRISPR-Cas12a and expanding the design space for programmable RNA manipulation.
External linksNat Biotechnol / PubMed:42067668
MethodsEM (single particle)
Resolution3.17 Å
Structure data

68245

22fn

EMDB-68245, PDB-22fn:
Cryo-EM structure of AsCas12a in complex with crDNA and RNA target
Method: EM (single particle) / Resolution: 3.17 Å

Source
  • acidaminococcus sp. bv3l6 (bacteria)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / Cas12a / crDNA / RNA

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