A photo-switchable gold nanoformulation based on the dCas9 protein for spatiotemporal controlled gene editing activation in vivo
Konstantinidou, S.; Rocco, M. A.; Nocilla, F.; Debellis, D.; d'Amora, M.; De Angelis, F.; Gabellini, C.; Fuso, F.; Tantussi, F.; Raffa, V.
Show abstract
Genome editing allows for the manipulation of genomic DNA for biotechnology and biomedical applications, but the specificity and control of the editing process remain a challenge. This study introduces a nano-switch for spatiotemporal control of the editing process. The basic module of the nano-switch (the monomer) is composed of a gold nanorod conjugated with the dead Cas9. Based on mathematical models, we established the design and the mechanism of action of the nano-switch. Briefly, when two monomers, guided by their respective guide RNAs, form a dimer onto the DNA and get irradiated with a Near-Infrared pulsed laser resonant at the plasmonic properties of the dimer, they generate a localized heat that triggers a thermal break onto the DNA. The nano-switch was generated, validated, and tested in zebrafish embryos at the 1-cell stage. Molecular analysis of irradiated embryos showed targeted DNA mutations, validating the efficacy of the nano-switch as a tool for conditional gene editing that integrates the if-when-where functions.
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