Light‐Triggered Genome Editing: Cre Recombinase Mediated Gene Editing with Near‐Infrared Light

A light‐activated genome editing platform based on the release of enzymes from a plasmonic nanoparticle carrier when exposed to biocompatible near‐infrared light pulses is described. The platform relies on the robust affinity of polyhistidine tags to nitrilotriacetic acid in the presence of copper which is attached to double‐stranded nucleic acids self‐assembled on the gold nanoparticle surface. A protein fusion of the Cre recombinase containing a TAT internalization peptide sequence to achieve endosomal localization is also employed. High‐resolution gene knock‐in of a red fluorescent reporter is observed using a commercial two‐photon microscope. High‐throughput irradiation is described to generate useful quantities of edited cells. Light‐controlled genome editing is demonstrated by delivering Cre recombinase, using plasmonic gold nanoparticles and 800 nm light. This allows for spatiotemporal controlled protein activity permitting future endeavors in precision genome editing.