CRISPR/Cas9技术在单纯疱疹病毒研究中的应用

Application of CRISPR/Cas9 technology in the study of herpes simplex virus

单纯疱疹病毒(herpes simplex virus,HSV)是导致人类疱疹性疾病的主要病原体,具有广泛宿主细胞类型、庞大外源基因容量等特点,因此也是一种颇具吸引力的病毒载体。HSV基因组较为庞大,对于病毒基因组的操作技术便成为关键环节。对该病毒基因组的操作先后经历了同源重组(homologous recombination),细菌人工染色体技术(bacterial artificial chromosome,BAC)和CRISPR/Cas9(clustered,regularly interspaced,short palindromic repeat/CRISPR-associated nuclease 9)技术。同源重组具有重组效率低、产物不易纯化等缺点,而且将大型病毒基因组克隆到BAC上是一个耗时耗力的过程。近年来,CRISPR/Cas9技术因其操作简便、重复性好、成本低廉等优势而被越来越多的研究者应用于各种物种的基因工程研究工作中,对于病毒基因组的操作也随之更加精确高效。本文对CRISPR/Cas9技术在HSV基因功能、治疗HSV感染性疾病及HSV载体研究中的应用进行综述,以期对相关的研究工作提供一定参考。

Herpes simplex virus(HSV),the main pathogen of human herpetic diseases,is an attractive viral vector due to a wide range of host cell types and a large capacity of foreign gene.The manipulation on herpes simplex virus genome,up to 152 kb,has been the key process and has undergone the improvement from homologous recombination to bacterial artificial chromosome(BAC)and CRISPR/Cas9.However,homologous recombination is imperfect because the efficiency of recombination is relatively low and the product is too complex to purify.Construction of recombinant HSV based on BAC is time-and labor-consuming.Currently,CRISPR/Cas9 has been applied to genetic engineering of various species by more and more researchers due to its convenience,good repeatability,low cost,and then the manipulation of viral genome is more accurate and efficient.Application of CRISPR/Cas9 in the research of HSV gene function,HSV infection therapy and viral vector were summarized in this review,in order to provide reference for corresponding research.