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.

可控制地和有效地定位的CRISPR / Cas9质粒转染在与各种人类关键疾病相关的基因编辑中起着至关重要的作用。我们采用了通过电荷逆转纳米载体传递近红外（NIR）光响应的CRISPR / Cas9质粒，以实现高效的位点特异性基因编辑。基于涂覆在上转换纳米材料（UCNP-UVP-P）上的紫外线敏感共轭聚电解质的基础上，制备了具有丰富正电荷的纳米载体，该载体可在980 nm光照射下转化为负电荷。使用制备的纳米载体，我们证明了该质粒可以以时间控制的方式有效地转染到肿瘤细胞（〜63％±4％）中，并且功能性CRISPR / Cas9蛋白可以在经选择的NIR照射下成功表达地区。将CRISPR / Cas9基因导入体内肿瘤细胞，在光照射下诱导目标基因PLK-1的高效编辑。因此，这种精确控制的基因调控策略有可能作为复杂生物系统中基因工程的新范例。