Abstract:
RNA interference (RNAi) serves as an indispensable tool for gene function studies and has been substantiated through extensive research for its practical applications in the baculovirus expression vector system (BEVS). This chapter expands the RNAi toolkit in insect cell culture by including small interfering RNA (siRNA) in the protocol, in addition to the conventional use of double-stranded RNA (dsRNA). This chapter also brings attention to key design and reporting considerations, based on Minimum Information About an RNAi Experiment (MIARE) guidelines. Recommendations regarding online tools for dsRNA and siRNA design are provided, along with guidance on choosing suitable methods for measuring silencing outcomes.
摘要:
RNA干扰(RNAi)是基因功能研究不可或缺的工具,并已通过广泛的研究证实了其在杆状病毒表达载体系统(BEVS)中的实际应用。本章通过在方案中包括小干扰RNA(siRNA)来扩展昆虫细胞培养中的RNAi工具包,除了常规使用双链RNA(dsRNA)。本章还提请注意关键的设计和报告考虑因素,基于关于RNAi实验(MIARE)指南的最低信息。提供了有关dsRNA和siRNA设计的在线工具的建议,以及选择合适的方法来测量沉默结果的指导。