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利用氟化α-螺旋多肽在体外和体内进行 U1 snRNA 干扰的 siRNA 介导的方案。

Protocol for siRNA-mediated U1 snRNA knockdown using fluorinated α-helical polypeptide in vitro and in vivo.

机构信息

National Key Laboratory of Immunity and Inflammation, and CAMS Key Laboratory of Synthetic Biology Regulatory Elements, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou 215123, China.

Institute of Functional Nano and Soft Materials (FUNSOM), Jiangsu Key Laboratory for Carbon-Based Functional Materials and Devices, Collaborative Innovation Center of Suzhou Nano Science and Technology, Soochow University, Suzhou 215123, China.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103238. doi: 10.1016/j.xpro.2024.103238. Epub 2024 Aug 2.

Abstract

Here, we present a protocol for small interfering RNA (siRNA)-mediated U1 small nuclear RNA (snRNA) knockdown using fluorinated α-helical polypeptide in macrophages and mouse lungs, providing a dependable approach to silence U1 snRNA in vitro and in vivo. We describe steps for preparing P7F7/siRNA polyplexes and silencing U1 snRNA with P7F7/siRNA polyplexes in macrophages and mouse lungs. Knockdown efficiency is validated through reverse-transcription quantitative real-time PCR analysis. This protocol is applicable for studying the physiological or pathophysiological function of U1 snRNA. For complete details on the use and execution of this protocol, please refer to Zhang et al..

摘要

在这里,我们提供了一种使用氟化 α-螺旋多肽在巨噬细胞和小鼠肺部中进行小干扰 RNA (siRNA) 介导的 U1 小核 RNA (snRNA) 敲低的方案,为在体外和体内沉默 U1 snRNA 提供了一种可靠的方法。我们描述了制备 P7F7/siRNA 复合物和用 P7F7/siRNA 复合物在巨噬细胞和小鼠肺部中沉默 U1 snRNA 的步骤。通过逆转录定量实时 PCR 分析验证敲低效率。该方案适用于研究 U1 snRNA 的生理或病理生理学功能。有关此方案使用和执行的完整详细信息,请参阅 Zhang 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1f56/11342768/2df69f2d52f3/fx1.jpg

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