9TQ4

Asp2Cas12l-sgRNA bound to target DNA

Summary for 9TQ4

• Entry DOI: 10.2210/pdb9tq4/pdb
• EMDB information: 56124
• Descriptor: Asp2Cas12l, sgRNA (149-MER), DNA (26-MER), ... (4 entities in total)
• Functional Keywords: crispr-cas, cas12, hydrolase
• Biological source: Armatimonadota; More
• Total number of polymer chains: 4
• Total formula weight: 181119.54

Authors

Sasnauskas, G.,Tamulaitiene, G.,Urbaitis, T.,Gasiunas, G. (deposition date: 2025-12-19, release date: 2026-08-19)

Primary citation

Urbaitis, T.,Trinkuniene, L.,Lenkaite, I.,Petrauskyte, M.,Krasauskas, R.,Stitilyte, M.,Sabaliauskas, M.,Sasnauskas, G.,Tamulaitiene, G.,Young, J.K.,Siksnys, V.,Gasiunas, G.
A Potent CRISPR-Cas12l Double-Strand Break Gene Editor.
CRISPR J, 9:126-140, 2026

PubMed Abstract: Recently, a new family of CRISPR-Cas12 endonucleases from an unexplored phylum of bacteria, , was discovered. Named Cas12l, they are compact (800-900 aa), recognize a 5' C-rich protospacer adjacent motif, and present an N-terminal domain that stretches from the beginning to the end of the ribonucleoprotein-bound DNA target site, effectively locking it in place. Here, structure-guided rational design supplemented with AI-based large protein language model predictions was used to improve rates of DNA target cleavage of a family member, Asp2Cas12l. Compared to the wild-type, engineered variants exhibited an approximately 10-fold increase in double-strand break (DSB) editing efficiency in human cells with less target-to-target variation. Moreover, frequencies of editing were comparable to those of SpCas9 at overlapping target sites, and their DSBs efficiently corrected by homology-directed repair (39-56% of editing outcomes). Altogether, this study extends our understanding of CRISPR-Cas12 protein engineering and offers a potent new alternative for DSB-mediated genome editing in human cells.

PubMed: 42163774
DOI: 10.1177/25731599261448428

Experimental method

ELECTRON MICROSCOPY (2.51 Å)