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RNAi 介导的昆虫细胞-杆状病毒表达系统沉默。

RNAi-Mediated Silencing in the Insect Cell-Baculovirus Expression System.

机构信息

Gene Therapy Franchise, National Resilience, Waltham, MA, USA.

出版信息

Methods Mol Biol. 2024;2829:91-107. doi: 10.1007/978-1-0716-3961-0_7.

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) 是研究基因功能不可或缺的工具,在杆状病毒表达载体系统 (BEVS) 中的实际应用也得到了广泛的研究证实。本章在昆虫细胞培养的 RNAi 工具包中增加了小干扰 RNA (siRNA),除了传统的双链 RNA (dsRNA) 的使用。本章还根据最小信息关于 RNAi 实验 (MIARE) 指南,关注了关键的设计和报告注意事项。此外,还提供了有关 dsRNA 和 siRNA 设计的在线工具的建议,并就如何选择合适的方法来衡量沉默效果提供了指导。

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