构建重组腺病毒-5载体，防止出现具有复制能力的腺病毒

IF 1.1 4区医学 Q4 VIROLOGY

Acta virologica Pub Date : 2023-12-19 DOI:10.3389/av.2023.11642

Wenbo Xie, Yifei Yuan, Bo Liu, Min Liang

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引用次数: 0

摘要

人腺病毒-5（hAd5）是一种重要的基因递送载体，已被广泛应用于基因治疗、癌症治疗和疫苗开发等生物医学的各个领域。然而，在 HEK293 细胞中制备腺病毒载体时产生的复制能力强的腺病毒（RCA）一直是一个令人担忧的问题。本研究对人腺病毒-5进行了改造，缩短了腺病毒与HEK293基因组DNA之间的同源序列长度，从而减少了RCA的产生。重组后的 hAd5 在 HEK293 细胞中扩增并连续传代 12 次。通过实时定量 PCR 检测第 2、4、6、8、10 和 12 次传代时 RCA 的含量。结果表明，修饰腺病毒载体可有效减少 RCA 在 HEK293 细胞中连续传代的产生。

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Construction of recombinant adenovirus-5 vector to prevent replication-competent adenovirus occurrence

Human adenovirus-5 (hAd5) is an important gene delivery vector, which has been widely used in various fields of biomedicine, such as gene therapy, cancer therapy, and vaccine development. However, replication-competent adenovirus (RCA) generated when adenoviral vectors are prepared in HEK293 cells has remained a concern. In this study, the human adenovirus-5 was modified to shorten the length of homologous sequence between the adenovirus and HEK293 genomic DNA, thereby reducing the production of RCA. The recombinant hAd5 was amplified and serially passaged 12 times in HEK293 cells. The amounts of RCA at passage 2, 4, 6, 8, 10, and 12 were detected by quantitative real-time PCR. The results demonstrated that the modification of adenoviral vector could effectively reduce the production of RCA during serial passages in HEK293 cells.

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来源期刊

Acta virologica 医学-病毒学

CiteScore

3.10

自引率

11.80%

发文量

审稿时长

>12 weeks

期刊介绍： Acta virologica is an international journal of predominantly molecular and cellular virology. Acta virologica aims to publish papers reporting original results of fundamental and applied research mainly on human, animal and plant viruses at cellular and molecular level. As a matter of tradition, also rickettsiae are included. Areas of interest are virus structure and morphology, molecular biology of virus-cell interactions, molecular genetics of viruses, pathogenesis of viral diseases, viral immunology, vaccines, antiviral drugs and viral diagnostics.