We herein report an optogenetically activatable CRISPR-Cas9 nanosystem for programmable genome editing in the second near-infrared (NIR-II) optical window. The nanosystem, termed nanoCRISPR, is composed of a cationic polymer-coated Au nanorod (APC) and Cas9 plasmid driven by a heat-inducible promoter. The APC not only serves as a carrier for intracellular plasmid delivery but also can harvest external NIR-II photonic energy and convert it into local heat to induce the gene expression of the Cas9 endonuclease. Due to high transfection activity, the APC shows strong ability to induce a significant level of disruption in different genomic loci upon optogenetic activation. Moreover, the precise control of genome-editing activity can be simply programmed by finely tuning exposure time and irradiation time in vitro and in vivo and also enables editing at multiple time points, thus proving the sensitivity and inducibility of such an editing modality. The NIR-II optical feature of nanoCRISPR enables therapeutic genome editing at deep tissue, by which treatment of deep tumor and rescue of fulminant hepatic failure are demonstrated as proof-of-concept therapeutic examples. Importantly, this modality of optogenetic genome editing can significantly minimize the off-target effect of CRISPR-Cas9 in most potential off-target sites. The optogenetically activatable CRISPR-Cas9 nanosystem we have developed offers a useful tool to expand the current applications of CRISPR-Cas9, and also defines a programmable genome-editing strategy toward high precision and spatial specificity.

中文翻译：

---

用于可编程基因组编辑的光热nanoCRISPR的近红外光遗传工程。

我们在此报告了在第二近红外（NIR-II）光学窗口中用于可编程基因组编辑的光遗传学可激活CRISPR-Cas9纳米系统。纳米系统被称为nanoCRISPR，由阳离子聚合物包被的Au纳米棒（APC）和由热诱导启动子驱动的Cas9质粒组成。APC不仅充当细胞内质粒递送的载体，而且可以收获外部NIR-II光子能并将其转化为局部热量以诱导Cas9核酸内切酶的基因表达。由于高转染活性，APC显示出强大的能力，能够在光遗传学激活后诱导不同基因组位点的显着水平的破坏。此外，通过微调体外和体内的暴露时间和照射时间，可以简单地编程基因组编辑活性的精确控制，并且还可以在多个时间点进行编辑，从而证明了这种编辑方式的敏感性和可诱导性。nanoCRISPR的NIR-II光学特性可在深部组织进行治疗性基因组编辑，从而证明了对深部肿瘤的治疗和暴发性肝衰竭的抢救是概念验证的治疗实例。重要的是，这种光遗传学基因组编辑的方式可以极大地降低CRISPR-Cas9在大多数潜在脱靶位点的脱靶作用。我们开发的可光遗传激活的CRISPR-Cas9纳米系统为扩展CRISPR-Cas9的当前应用提供了有用的工具，