9KWC

Cas12a-PCPS-light

Summary for 9KWC

• Entry DOI: 10.2210/pdb9kwc/pdb
• EMDB information: 62607
• Descriptor: LbCas12a, RNA (25-MER) (2 entities in total)
• Functional Keywords: cas12a-pcps-dark, dna binding protein/rna, header dna binding protein-rna complex, header dna binding protein/rna
• Biological source: Lachnospiraceae bacterium ND2006; More
• Total number of polymer chains: 2
• Total formula weight: 151531.64

Authors

Zhang, M.F. (deposition date: 2024-12-05, release date: 2025-07-16)

Primary citation

Hu, M.,Zhang, B.,Shan, Y.,Cao, F.,Wang, Y.,Qi, W.,Wang, X.,Shen, Y.,Guo, X.,Zhang, M.,Tian, T.,Xie, W.,Zhang, M.,Liang, F.,Pei, D.,Zhou, X.
Scalable modulation of CRISPR‒Cas enzyme activity using photocleavable phosphorothioate DNA.
Nat Commun, 16:5939-5939, 2025

PubMed Abstract: The regulation of CRISPR‒Cas activity is critical for developing advanced biotechnologies. Optical control of CRISPR‒Cas system activity can be achieved by modulation of Cas proteins or guide RNA (gRNA), but these approaches either require complex protein engineering modifications or customization of the optically modulated gRNAs according to the target. Here, we present a method, termed photocleavable phosphorothioate DNA (PC&PS DNA)-mediated regulation of CRISPR‒Cas activity (DNACas), that is versatile and overcomes the limitations of conventional methods. In DNACas, CRISPR‒Cas activity is silenced by the affinity binding of PC&PS DNA and restored through light-triggered chemical bond breakage of PC&PS DNA. The universality of DNACas is demonstrated by adopting the PC&PS DNA to regulate various CRISPR‒Cas enzymes, achieving robust light-switching performance. DNACas is further adopted to develop a light-controlled one-pot LAMP-BrCas12b detection method and a spatiotemporal gene editing strategy. We anticipate that DNACas could be employed to drive various biotechnological advances.

PubMed: 40593724
DOI: 10.1038/s41467-025-61094-5

Experimental method

ELECTRON MICROSCOPY (2.9 Å)