根据转染的 dsRNA 确定 Dicer 作为 dsRNA 结合和裂解试剂的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-24 DOI:10.1016/j.xpro.2024.103591

Yunpeng Dai, Jiaxin Wang, Jiaqi Zhang, Xing Liu, Gang Sun, Jinfeng Lu, Yang Li

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引用次数: 0

摘要

哺乳动物Dicer已被证明对双链rna （dsRNAs）具有功能，并参与抗病毒免疫或免疫调节。在这里，我们提出了一种基于小RNA测序（RNA-seq）和dsRNA免疫沉淀（dsRNA- ip）的方案，用于鉴定Dicer是细胞系中转染dsRNA的dsRNA结合和切割因子。我们详细介绍了小RNA-seq数据的实验过程和分析。该协议可以进一步应用于鉴定将dsRNA加工成小rna的蛋白质，如RNase L.有关该协议使用和执行的完整细节，请参阅Dai等人1。

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Protocol for identifying Dicer as dsRNA binding and cleaving reagent in response to transfected dsRNA.

Mammalian Dicer has been proved to be functional on double-stranded RNAs (dsRNAs) and involved in antiviral immunity or immune regulation. Here, we present a protocol for identifying Dicer as a dsRNA binding and cleaving factor to transfected dsRNA in cell lines, based on small RNA sequencing (RNA-seq) and dsRNA-immunoprecipitation (dsRNA-IP). We detail both experimental processes and analysis on small RNA-seq data. This protocol can be further applied to identify proteins that process dsRNA into small RNAs, like RNase L. For complete details on the use and execution of this protocol, please refer to Dai et al.¹.

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