| Title |
A potent CRISPR-Cas12l double-strand break gene editor |
| Authors |
Urbaitis, Tomas ; Trinkūnienė, Laima ; Lenkaitė, Ieva ; Petrauskytė, Monika ; Krasauskas, Renatas ; Štitilytė, Miglė ; Sabaliauskas, Modestas ; Sasnauskas, Giedrius ; Tamulaitienė, Giedrė ; Young, Joshua K ; Šikšnys, Virginijus ; Gasiūnas, Giedrius |
| DOI |
10.1177/25731599261448428 |
| Full Text |
|
| Is Part of |
CRISPR journal.. Thousand Oaks : SAGE Publications. 2026, Early Access, p. [1-15].. ISSN 2573-1599. eISSN 2573-1602 |
| Keywords [eng] |
CRISPR-Cas12l ; double-strand break ; Asp2Cas12l |
| Abstract [eng] |
Recently, a new family of CRISPR-Cas12 endonucleases from an unexplored phylum of bacteria, Armatimonadota , 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. |
| Published |
Thousand Oaks : SAGE Publications |
| Type |
Journal article |
| Language |
English |
| Publication date |
2026 |
| CC license |
|
