Near-infrared optogenetic engineering of photothermal nanoCRISPR for programmable genome editing

Significance The persistent Cas9 activity following on-target editing and the lack of spatial precision have severely constrained the current CRISPR-Cas9 systems from complicated and diverse genome-editing contexts. Herein, we develop an optogenetically activatable CRISPR-Cas9 nanosystem (termed nanoCRISPR), and demonstrate that programmable and inducible genome editing can be simply manipulated through photothermal regulation of nanoCRISPR in the second near-infrared optical window. The genome-editing activity and spatial specificity can be precisely programmed by photothermal activation of nanoCRISPR in vitro and in vivo, and the optogenetic regulation minimizes the off-target mutations of CRISPR-Cas9 in potential off-target sites. The photothermal nanoCRISPR offers a useful tool to expand the current applications of CRISPR-Cas9 toward high precision and spatial specificity.