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 catalytically 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.
A photo-inducible nano-switch for spatiotemporal controlled gene editing activation
Konstantinidou, Soultana;d'Amora, Marta;Nocilla, Francesco;Gabellini, Chiara;Fuso, Francesco;Raffa, Vittoria
2025-01-01
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 catalytically 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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
