Wang Jiaming, Dai Wenxin, Zhou Shuqi, Ma Wenfu
School of Life Sciences, Beijing University of Chinese Medicine.
School of Life Sciences, Beijing University of Chinese Medicine;
J Vis Exp. 2025 Jun 6(220). doi: 10.3791/68389.
The severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) virus-like particle (SC2-VLP) method offers a powerful and accessible tool for studying the SARS-CoV-2 life cycle without the need for biosafety level 3 (BSL-3) laboratories. This system effectively mimics critical stages of the viral life cycle, including assembly, genome packaging, and egress, using a luciferase reporter fused to the T20 signal for sensitive and precise detection of viral particle production. SC2-VLPs are generated by co-expressing SARS-CoV-2 structural proteins, including membrane (M), nucleocapsid (N), envelop (E), and spike (S), along with the RNA packaging signal in HEK-293T cells. Unlike traditional virus-like particle systems, the SC2-VLP method ensures accurate quantification and greater fidelity to the natural viral life cycle. Furthermore, compared to lentiviral pseudotyping methods, which are limited to studying viral entry through the incorporation of S protein into HIV-based lentiviral particles, the SC2-VLP system provides a more comprehensive platform for exploring multiple stages of SARS-CoV-2 biology. While this method bypasses the risks of handling live virus and expands accessibility. The SC2-VLP method represents a significant advancement in antiviral research and the development of therapeutic strategies against SARS-CoV-2.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）病毒样颗粒（SC2-VLP）方法提供了一种强大且易于使用的工具，用于研究SARS-CoV-2的生命周期，而无需生物安全3级（BSL-3）实验室。该系统使用与T20信号融合的荧光素酶报告基因，有效模拟病毒生命周期的关键阶段，包括组装、基因组包装和释放，以灵敏且精确地检测病毒颗粒的产生。SC2-VLPs是通过在HEK-293T细胞中共表达SARS-CoV-2结构蛋白（包括膜蛋白（M）、核衣壳蛋白（N）、包膜蛋白（E）和刺突蛋白（S））以及RNA包装信号而产生的。与传统的病毒样颗粒系统不同，SC2-VLP方法可确保准确量化并更忠实地模拟天然病毒生命周期。此外，与慢病毒假型化方法相比，慢病毒假型化方法仅限于通过将S蛋白掺入基于HIV的慢病毒颗粒中来研究病毒进入，而SC2-VLP系统为探索SARS-CoV-2生物学的多个阶段提供了更全面的平台。虽然这种方法规避了处理活病毒的风险并扩大了可及性，但SC2-VLP方法代表了抗病毒研究以及针对SARS-CoV-2治疗策略开发方面的一项重大进展。
