TitanCas12h

TitanCas12h is an engineered Type V-H CRISPR-Cas12h nuclease developed by Reforgene Medicine for eukaryotic genome editing. Discovered through metagenomic mining using protein language models and optimized via AI-driven protein engineering and rational design (specifically variant Mut-14), TitanCas12h overcomes the natural limitation of wild-type Cas12h effectors, which typically function as nickases. It is capable of inducing efficient double-strand breaks (DSBs) with a unique deletion profile (15–26 bp), making it particularly effective for gene disruption. The system recognizes a WYR PAM and has demonstrated high specificity with minimal off-target activity. Preclinical studies have validated its efficacy in vivo using lipid nanoparticle (LNP) delivery of mRNA and gRNA to target cardiovascular-related genes such as ANGPTL3, AGT, LPA, and LDLR, showing significant protein depletion in humanized mouse models.