Wu Yiyi, Gao Shangqing, Liu Guanya, Wang Mengwei, Tan Ruixiao, Huang Baoying, Tan Wenjie
National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, 155 Changbai Road, Beijing, 102206, China.
School of Public Health, Baotou Medical College, Baotou City, Inner Mongolia Autonomous Region, 014040, China.
Mol Biomed. 2025 Apr 29;6(1):26. doi: 10.1186/s43556-025-00266-7.
Infectious Clones represent a foundational technique in the field of reverse genetics, allowing for the construction and manipulation of full-length viral genomes. The main methods currently used for constructing viral infectious clones include Transformation-associated recombination (TAR), which is based on Yeast Artificial Chromosome (YAC) and Bacterial Artificial Chromosome (BAC). The YAC and BAC systems are powerful tools that enable the clones and manipulation of large DNA fragments, making them well-suited for the construction of full-length viral genomes. These methods have been successfully applied to construct infectious clones for a wide range of viruses, including coronaviruses, herpesviruses, flaviviruses and baculoviruses. The rescued recombinant viruses from these infectious clones have been widely used in various research areas, such as vaccine development, antiviral drug screening, pathogenesis and virulence studies, gene therapy and vector design. However, as different viruses possess unique biological characteristics, the challenge remains in how to rapidly obtain infectious clones for future research. In summary, this review introduced the development and applications of infectious clones, with a focus on the YAC, BAC and combined YAC-BAC technologies. We emphasize the importance of these platforms in various research areas and aim to provide deeper insights that can advance the platform and broaden its application horizons.
感染性克隆是反向遗传学领域的一项基础技术,可用于构建和操作全长病毒基因组。目前用于构建病毒感染性克隆的主要方法包括基于酵母人工染色体(YAC)和细菌人工染色体(BAC)的转化相关重组(TAR)。YAC和BAC系统是强大的工具,能够克隆和操作大片段DNA,非常适合用于构建全长病毒基因组。这些方法已成功应用于构建多种病毒的感染性克隆,包括冠状病毒、疱疹病毒、黄病毒和杆状病毒。从这些感染性克隆中拯救出的重组病毒已广泛应用于各个研究领域,如疫苗开发、抗病毒药物筛选、发病机制和毒力研究、基因治疗和载体设计。然而,由于不同病毒具有独特的生物学特性,如何快速获得用于未来研究的感染性克隆仍然是一个挑战。总之,本综述介绍了感染性克隆的发展和应用,重点介绍了YAC、BAC以及YAC-BAC联合技术。我们强调这些平台在各个研究领域的重要性,旨在提供更深入的见解,以推动该平台的发展并拓宽其应用前景。