TIGRa: An ultra-compact programmable activator enabling multiplexed and efficient gene regulation

Abstract

SUMMARY Programmable gene activation has broad therapeutic potential but remains constrained by large effector size, limited multiplexing capacity, and challenges in in vivo delivery. Here, we develop TIGRa, a compact transcriptional activator derived from the TIGR-TasR system that is mechanistically distinct from CRISPR-based activators. TIGRa is less than half the size of dSpCas9-based activators while achieving comparable or greater activation efficiency. Its native TIGR array architecture enables efficient multiplexed regulation, supporting simultaneous activation of up to 12 endogenous genes from a single compact construct. TIGRa-mediated multi-gene activation efficiently reprogrammed human fibroblasts into induced pluripotent stem cells. In addition, an all-in-one AAV-TIGRa vector activated endogenous CaMKII in vivo, promoting retinal ganglion cell survival and preserving visual function in a mouse model of NMDA-induced retinal injury. These results establish TIGRa as a compact and multiplexable platform for therapeutic gene regulation and in vivo genetic medicine.