新型基因编辑技术CRISPR/Cas9的出现，为精准化基因治疗开启了新的篇章，但目前仍停留在细胞水平。如何将CRISPR/Cas9向CNS有效投递是靶向基因治疗的关键。基于超声介导微泡靶向开放BBB，课题组成功构建多种跨BBB和靶向性载药或载基因体系。本项目提出设计一载sgRNA/Cas9的纳米级、多功能基因组学编辑新型载体，通过级联靶向策略，实现针对胶质瘤的精准治疗。该系统以靶向核酸适配体修饰脂质体，包载靶向敲除MGMT基因的sgRNA/Cas9复合物，以声致相变型PLGA液态氟碳纳米粒为系统核心。在超声介导下相变开放BBB，借助核酸适配体与胶质瘤特异性结合，促进内化，通过下调MGMT表达，增强胶质瘤对化疗的敏感性。所建纳米系统经物理表征、包载率、毒性、离体和载体动物靶向性等试验验证，实现CRISPR/Cas9在CNS的精准投递和基因编辑。为探索CNS疾病的精准化基因治疗提供新的途径。

The emergence of CRISPR/Cas9, a new kind of genome-editing technique, opened a new chapter on precise gene therapy. But most of the studies still remained at cellular level recently. The high efficiency delivery of CRISPR/Cas9 to central nervous system is the key for targeted gene therapy of CNS disease. Our group had constructed several targeted gene or drug loaded systems which could across blood brain barrier under ultrasound combined with microbubbles. This project proposed the design of a new vector of sgRNA/Cas9 for nanoscale, multifunctional genomics editing, and achieved a precise treatment for glioma through cascade targeting strategy. The liposome in this system is modified with aptamer, and the sgRNA/Cas9 complex is designed to knock out the MGMT gene, and the acoustic phase change PLGA-liquid fluorocarbon nanoparticle is the core of the system. Ultrasound mediated phase change could open BBB, and the specific binding between aptamer and glioma could promote internalization of sgRNA/Cas9-loaded liposome, leading to down regulation of MGMT expression and enhancement of the sensitivity of glioma on chemotherapy. This nano system will be verified by physical characterization, loading rate, toxicity, targeting efficiency in vitro and in vivo, for achieving accurate delivery and gene editing of CRISPR / Cas9 in CNS. To provide a new way to explore the precise gene therapy for CNS diseases.