Rotaviruses (RVs) are considered to be one of the most common causes of viral gastroenteritis in young children and infants worldwide. Before recent developments, studies on rotavirus biology have suffered from the lack of an effective reverse genetics (RG) system to generate recombinant rotaviruses and study the precise roles of the viral proteins in the context of RV infection. Lately a fully-tractable plasmid-only based RG system for rescuing recombinant rotaviruses has been developed leading to a breakthrough in the RV field. Since then, the reproducibility and improvements of this technology have led to the generation of several recombinant rotaviruses with modifications on different gene segments, which has allowed the manipulation of viral genes to characterise the precise roles of viral proteins during RV replication cycle or to encode exogenous proteins for different purposes.

This review will recapitulate the different RG approaches developed so far, highlighting any similarities, differences and limitations of the systems as well as the gene segments involved. The review will further summarise the latest recombinant rotaviruses generated using the plasmid-only based RG system showing the enormous potentials of this technique to shed light on the still unanswered questions in rotavirus biology.

轮状病毒 (RVs) 被认为是全球幼儿和婴儿病毒性胃肠炎的最常见原因之一。在最近的发展之前，轮状病毒生物学研究一直缺乏有效的反向遗传学 (RG) 系统来产生重组轮状病毒并研究病毒蛋白在 RV 感染中的确切作用。最近开发了一种完全可处理的仅基于质粒的 RG 系统，用于拯救重组轮状病毒，从而在 RV 领域取得了突破。从那时起，该技术的可重复性和改进导致产生了几种重组轮状病毒，这些病毒对不同的基因片段进行了修饰，

这篇综述将概括迄今为止开发的不同 RG 方法，强调系统以及所涉及的基因片段的任何相似性、差异和局限性。该综述将进一步总结使用仅基于质粒的 RG 系统生成的最新重组轮状病毒，显示该技术在阐明轮状病毒生物学中仍未解决的问题方面的巨大潜力。