22FN

Cryo-EM structure of AsCas12a in complex with crDNA and RNA target

Summary for 22FN

• Entry DOI: 10.2210/pdb22fn/pdb
• EMDB information: 68245
• Descriptor: RNA (5'-R(P*GP*AP*CP*AP*GP*CP*CP*CP*AP*CP*AP*UP*GP*GP*CP*AP*UP*UP*CP*CP*AP*CP*U)-3'), CRISPR-associated endonuclease Cas12a, DNA (41-MER) (3 entities in total)
• Functional Keywords: cas12a, crdna, rna, dna binding protein
• Biological source: Acidaminococcus sp. BV3L6; More
• Total number of polymer chains: 3
• Total formula weight: 178942.97

Authors

Lam, W.H.,Wu, X.,Hsing, I.M.,Zhai, Y. (deposition date: 2026-01-08, release date: 2026-04-29, Last modification date: 2026-05-13)

Primary citation

Wu, X.,Lam, W.H.,Zhao, Z.,Cao, Y.,Lin, H.,Feng, X.,Zhai, Y.,Hsing, I.M.
DNA-guided CRISPR-Cas12a effectors for programmable RNA recognition and cleavage.
Nat.Biotechnol., 2026

PubMed Abstract: 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.

PubMed: 42067668
DOI: 10.1038/s41587-026-03120-5

Experimental method

ELECTRON MICROSCOPY (3.17 Å)