Asp2Cas12l

Asp2Cas12l is a compact CRISPR-Cas12l endonuclease, typically ranging from 800 to 900 amino acids, discovered within the bacterial phylum Armatimonadota. It is characterized by its recognition of a 5' C-rich protospacer adjacent motif (PAM) and a unique N-terminal domain that stabilizes the ribonucleoprotein (RNP)-bound DNA target site. Engineered variants of Asp2Cas12l, developed through structure-guided rational design and AI-based protein language model predictions, exhibit significantly enhanced double-strand break (DSB) editing efficiency in human cells, comparable to Streptococcus pyogenes Cas9 (SpCas9). The enzyme maintains high specificity and has been evaluated for therapeutic potential using adeno-associated virus (AAV) delivery via subretinal injection in murine models, suggesting potential applications in gene therapy for ocular and other genetic disorders.