在悬浮细胞系中通过多核糖体谱分析和定量PCR来鉴定潜在的编码微小肽的长链非编码RNA。

Han Cai, Sun Linyu, Pan Qi, Sun Yumeng, Wang Wentao, Chen Yueqin

MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong 510275, China.

STAR Protoc. 2021 Dec 14;3(1):101037. doi: 10.1016/j.xpro.2021.101037. eCollection 2022 Mar 18.

Micropeptides are emerging as important regulators of various cellular processes. Long non-coding RNAs (lncRNAs) serve as a source of micropeptide-encoding small reading frames. The techniques to detect micropeptides or translating lncRNAs, such as mass spectrometry and ribosome profiling, are sophisticated and expensive. Here, we present an easy and cost-effective protocol to screen for potential micropeptide-encoding lncRNAs by polysome profiling in suspension cell lines. When combined with quantitative PCR, this protocol facilitates the identification of a number of translating lncRNAs simultaneously. For complete details on the use and execution of this protocol, please refer to Sun et al. (2021).

微小肽正逐渐成为各种细胞过程的重要调节因子。长链非编码RNA（lncRNA）作为微小肽编码小阅读框的来源。检测微小肽或正在翻译的lncRNA的技术，如质谱分析和核糖体分析，复杂且昂贵。在此，我们提出了一种简单且经济高效的方案，通过悬浮细胞系中的多核糖体分析来筛选潜在的微小肽编码lncRNA。当与定量PCR结合使用时，该方案有助于同时鉴定多个正在翻译的lncRNA。有关此方案的使用和执行的完整详细信息，请参考Sun等人（2021年）的文献。