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

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 efficiently transfected into tumor cells (∼ 63% ± 4%) in a time-controlled 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 photoirradiation. Therefore, this precisely controlled gene regulation strategy has the potential to serve as a new paradigm for gene engineering in complex biological systems.