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

光触发基因组编辑:利用近红外光进行Cre重组酶介导的基因编辑

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作者:Demosthenes P Morales,Erin N Morgan,Meghan McAdams,Amanda B Chron,Jeong Eun Shin,Joseph A Zasadzinski,Norbert O Reich

Abstract

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.

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