Abstract：
The adeno-associated virus (AAV) is a defective single-stranded DNA virus with the simplest structure reported to date. It constitutes a capsid protein and single-stranded DNA. With its high transduction efficiency, low immunogenicity, and tissue specificity, it is the most widely used and promising gene therapy vector. The clustered regularly interspaced short palindromic sequence (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing system is an emerging technology that utilizes cas9 nuclease to specifically recognize and cleave target genes under the guidance of small guide RNA and realizes gene editing through homologous directional repair and non-homologous recombination repair. In recent years, an increasing number of animal experiments and clinical studies have revealed the great potential of AAV as a vector to deliver the CRISPR/cas9 system for treating genetic diseases and viral infections. However, the immunogenicity, toxicity, low transmission efficiency in brain and ear tissues, packaging size limitations of AAV, and immunogenicity and off-target effects of Cas9 protein pose several clinical challenges. This research reviews the role, challenges, and countermeasures of the AAV-CRISPR/cas9 system in gene therapy.
摘要：
腺相关病毒（AAV）是一种有缺陷的单链DNA病毒，具有迄今为止报道的最简单的结构。它构成衣壳蛋白和单链DNA。凭借其高转导效率，低免疫原性,和组织特异性，它是最广泛使用和最有前途的基因治疗载体。成簇规则间隔短回文序列(CRISPR)/CRISPR相关蛋白9(Cas9)基因编辑系统是在小向导RNA指导下,利用Cas9核酸酶特异性识别和切割靶基因,通过同源定向修复和非同源重组修复实现基因编辑的新兴技术。近年来,越来越多的动物实验和临床研究揭示了AAV作为载体提供CRISPR/cas9系统用于治疗遗传疾病和病毒感染的巨大潜力。然而,免疫原性，毒性,大脑和耳朵组织的低传输效率，AAV的包装尺寸限制，Cas9蛋白的免疫原性和脱靶效应构成了一些临床挑战。这项研究回顾了作用，挑战,AAV-CRISPR/cas9系统在基因治疗中的应用及对策。
