A smaller engineered Cas12m protein for activating multiple genes at once in living organisms

A single all-in-one AAV vector can deliver up to eight crRNAs for the activation of six different genes.

• The Cas12m protein family was engineered to create an optimized variant, denCas12m, for CRISPR-based regulation.
• This system allows for the processing of a multi-gRNA array into individual functional guides within target cells.
• Efficient activation of a single gene can occur through multiple sites, or distinct genes can be activated simultaneously.
• Phenotypic rescue was observed in mouse models of Duchenne muscular dystrophy and Scn1a haploinsufficiency.
• The platform may enable high-order multiplexing for gene activation using a single vector.

Simplified