An improved reverse genetics system for rotavirus vaccine strain LLR using five plasmid vectors.

Species A rotaviruses (RVs), which belong to the family Sedoreoviridae and contain a genome of 11 segmented dsRNA segments, are a leading cause of severe acute gastroenteritis in infants and children younger than 5 years of age. We previously developed a strategy to recover rotavirus vaccine strain LLR from 11 cloned plasmids. Here, we report an improved reverse genetics system for LLR by combining two or three transcriptional cassettes in a single plasmid, which substantially enhances rescue efficiency from 66.7% (8/12) to 91.7 % (11/12). Furthermore, the recombinant LLR stably expressing NLuc was rescued based on the five-plasmid reverse genetics system. Improvements to the rotavirus reverse genetics system will enhance its applicability for studies of rotavirus biology and clinical use.

Citation

Xiafei Liu, Junjie Yu, Shan Li, Pengdi Chai, Zhiping Xie, Lili Pang, Jinsong Li, Wuyang Zhu, Weihong Ren, Zhaojun Duan. An improved reverse genetics system for rotavirus vaccine strain LLR using five plasmid vectors. The Journal of general virology. 2025 Jan;106(1)

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PMID: 39791472

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