Light-responsive charge-reversal nanovector for high-efficiency in vivo CRISPR/Cas9 gene editing with controllable location and time

摘要

Controllably and efficaciously localized CRISPR/Cas9 plasmids transfection plays an essential role in genetic editing associated with various key human diseases.We employed near-infrared(NIR)light-responsive CRISPR/Cas9 plasmids delivery via a charge-reversal nanovector to achieve highly efficient and site-specific gene editing.The nanovector with abundant positive charges was fabricated on the basis of an ultraviolet-sensitive conjugated polyelectrolyte coated on an upconversion nanomaterial(UCNP-UVP-P),which can convert into negative charges upon 980 nm light irradiation.Using the as-prepared nanovector,we demonstrated the plasmids could be efficiency transfected into tumor cells(~63%±4%)in a time-contolled manner,and that functional CRISPR/Cas9 proteins could be successfully expressed in a selected NIR-irradiated region.Particularly,this strategy was successfully applied to the delivery of CRISPR/Cas9 gene to tumor cells in vivo,inducing high efficiency editing of the target gene PLK-1 under photolrradiation.Therefore,this precisely controlled gene regulation strategy has the potential to serve as a new paradigm for gene engineering in complex biological systems.