Zhang Nianchao, Li Jie, Yu Jingxuan, Wan Yajuan, Zhang Cuizhu, Zhang Hongkai, Cao Youjia
College of Life Sciences, Key Laboratory of Microbial Functional Genomics of the Ministry of Education, Nankai University, 94 Weijin Road, Tianjin 300071, China.
State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, China.
Virus Res. 2023 Jan 2;323:198979. doi: 10.1016/j.virusres.2022.198979. Epub 2022 Oct 23.
Oncolytic viruses are an emerging cancer treatment modality with promising results in clinical trials. The new generation of oncolytic viruses are genetically modified to enhance virus selectivity for tumor cells and allow local expression of therapeutic genes in tumors. The traditional technique for viral genome engineering based on homologous recombination using a bacterial artificial chromosome (BAC) system is laborious and time-consuming. With the advent of the CRISPR/Cas9 system, the efficiency of gene editing in human cells and other organisms has dramatically increased. In this report, we successfully applied the CRISPR/Cas9 technique to construct an HSV-based oncolytic virus, where the ICP34.5 coding region was replaced with the therapeutic genes murine interleukin 12 (IL12, p40-p35) and C-X-C motif chemokine ligand 11 (CXCL11), and ICP47 gene was deleted. The combination of IL12 and CXCL11 in oncolytic viruses showed considerable promise in colorectal cancer (CRC) treatment. Overall, our study describes genetic modification of the HSV-1 genome using the CRISPR/Cas9 system and provides evidence from principle studies for engineering of the HSV genome to express foreign genes.
溶瘤病毒是一种新兴的癌症治疗方式，在临床试验中取得了令人鼓舞的结果。新一代溶瘤病毒经过基因改造，以增强病毒对肿瘤细胞的选择性，并使治疗性基因在肿瘤中局部表达。基于使用细菌人工染色体（BAC）系统进行同源重组的传统病毒基因组工程技术既费力又耗时。随着CRISPR/Cas9系统的出现，人类细胞和其他生物体中的基因编辑效率大幅提高。在本报告中，我们成功应用CRISPR/Cas9技术构建了一种基于单纯疱疹病毒（HSV）的溶瘤病毒，其中ICP34.5编码区被治疗性基因小鼠白细胞介素12（IL12，p40-p35）和C-X-C基序趋化因子配体11（CXCL11）取代，并且ICP47基因被删除。溶瘤病毒中IL12和CXCL11的组合在结直肠癌（CRC）治疗中显示出相当大的前景。总体而言，我们的研究描述了使用CRISPR/Cas9系统对HSV-1基因组进行基因改造，并为改造HSV基因组以表达外源基因的原理研究提供了证据。
