State Key Laboratory of Virology, RNA Institute, College of Life Sciences and Frontier Science Center for Immunology and Metabolism, Wuhan University, Wuhan, 430072, China.
Institute for Vaccine Research at Animal Bio-safety Level Ⅲ Laboratory, Wuhan University School of Medicine, Wuhan, 430071, China.
Virol Sin. 2024 Jun;39(3):447-458. doi: 10.1016/j.virs.2024.03.009. Epub 2024 Mar 26.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still epidemic around the world. The manipulation of SARS-CoV-2 is restricted to biosafety level 3 laboratories (BSL-3). In this study, we developed a SARS-CoV-2 ΔN-GFP-HiBiT replicon delivery particles (RDPs) encoding a dual reporter gene, GFP-HiBiT, capable of producing both GFP signal and luciferase activities. Through optimal selection of the reporter gene, GFP-HiBiT demonstrated superior stability and convenience for antiviral evaluation. Additionally, we established a RDP infection mouse model by delivering the N gene into K18-hACE2 KI mouse through lentivirus. This mouse model supports RDP replication and can be utilized for in vivo antiviral evaluations. In summary, the RDP system serves as a valuable tool for efficient antiviral screening and studying the gene function of SARS-CoV-2. Importantly, this system can be manipulated in BSL-2 laboratories, decreasing the threshold of experimental requirements.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）仍在全球流行。SARS-CoV-2 的操作仅限于生物安全级别 3 实验室（BSL-3）。在这项研究中，我们开发了一种编码双报告基因 GFP-HiBiT 的 SARS-CoV-2 ΔN-GFP-HiBiT 复制子传递颗粒（RDP），能够产生 GFP 信号和荧光素酶活性。通过对报告基因的优化选择，GFP-HiBiT 在抗病毒评估中表现出更高的稳定性和便利性。此外，我们通过慢病毒将 N 基因递送至 K18-hACE2 KI 小鼠，建立了 RDP 感染小鼠模型。该小鼠模型支持 RDP 复制，可用于体内抗病毒评估。总之，RDP 系统是一种高效的抗病毒筛选和研究 SARS-CoV-2 基因功能的有价值的工具。重要的是，该系统可以在 BSL-2 实验室中操作，降低了实验要求的门槛。
