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TitleA Potent CRISPR-Cas12l Double-Strand Break Gene Editor.
Journal, issue, pagesCRISPR J, Vol. 9, Issue 3, Page 126-140, Year 2026
Publish dateMay 21, 2026
AuthorsTomas Urbaitis / Laima Trinkuniene / Ieva Lenkaite / Monika Petrauskyte / Renatas Krasauskas / Migle Stitilyte / Modestas Sabaliauskas / Giedrius Sasnauskas / Giedre Tamulaitiene / Joshua K Young / Virginijus Siksnys / Giedrius Gasiunas /
PubMed AbstractRecently, 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 ...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.
External linksCRISPR J / PubMed:42163774
MethodsEM (single particle)
Resolution2.51 Å
Structure data

56124

9tq4

EMDB-56124, PDB-9tq4:
Asp2Cas12l-sgRNA bound to target DNA
Method: EM (single particle) / Resolution: 2.51 Å

Source
  • armatimonadota (bacteria)
  • synthetic construct (others)
KeywordsHYDROLASE / CRISPR-Cas / Cas12

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