First Versatile Reverse Genetics System for DNA Viruses Using Circular Polymerase Extension Reaction.

IF 1.8 4区医学 Q4 IMMUNOLOGY

Microbiology and Immunology Pub Date : 2025-10-12 DOI:10.1111/1348-0421.70016

Hirotaka Yamamoto, Tomokazu Tamura, Rigel Suzuki, Saori Suzuki, Takasuke Fukuhara

引用次数: 0

Abstract

Reverse genetics enables the generation of recombinant viruses; however, conventional approaches using full-length cDNA cloned into bacterial artificial chromosomes or plasmids are time-consuming and technically challenging. While the circular polymerase extension reaction (CPER) has been applied to RNA viruses, it has not been used for DNA viruses. Here, we successfully applied CPER to generate infectious recombinant adenoviruses (AdVs) of two serotypes. The resulting viruses replicated comparably to parental strains, and replication was confirmed by immunostaining. These findings demonstrate that CPER is a rapid and efficient platform for reverse genetics of DNA viruses and may accelerate AdV research and therapeutic development.

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首个使用环状聚合酶延伸反应的DNA病毒多功能反向遗传系统。

反向遗传可以产生重组病毒；然而，将全长cDNA克隆到细菌人工染色体或质粒中的传统方法既耗时又具有技术挑战性。虽然环状聚合酶延伸反应（CPER）已应用于RNA病毒，但尚未用于DNA病毒。在这里，我们成功地将CPER应用于两种血清型的感染性重组腺病毒（AdVs）。由此产生的病毒复制与亲本株相当，并通过免疫染色证实了复制。这些发现表明，CPER是一个快速有效的DNA病毒反向遗传学平台，可能加速AdV的研究和治疗开发。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microbiology and Immunology 医学-免疫学

CiteScore

5.20

自引率

3.80%

发文量

审稿时长

1 months

期刊介绍： Microbiology and Immunology is published in association with Japanese Society for Bacteriology, Japanese Society for Virology, and Japanese Society for Host Defense Research. It is peer-reviewed publication that provides insight into the study of microbes and the host immune, biological and physiological responses. Fields covered by Microbiology and Immunology include:Bacteriology|Virology|Immunology|pathogenic infections in human, animals and plants|pathogenicity and virulence factors such as microbial toxins and cell-surface components|factors involved in host defense, inflammation, development of vaccines|antimicrobial agents and drug resistance of microbes|genomics and proteomics.