Integrated workflow for discovery of microprotein-coding small open reading frames.

Integrated workflow for discovery of microprotein-coding small open reading frames.
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DOI:
10.1016/j.xpro.2023.102649
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发表时间:
2023-12-15
期刊:
影响因子:
--
通讯作者:
Martinez, Thomas Farid
Martinez, Thomas Farid
中科院分区:
其他
文献类型:
--
作者:
Cao, Kevin;Heydary, Yasamin Hajy;Tong, Gregory;Martinez, Thomas Farid

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小开放阅读框(smORF)编码的微蛋白是一类含有少于100-150个氨基酸的新型功能性生物分子。在这里,我们提出了一种在哺乳动物系统全基因组范围内鉴定翻译smorf的方案。我们描述了生成核糖体分析(Ribo-seq)数据的步骤,转录组组装的计算机翻译以创建ORF数据库,以及Ribo-seq的计算分析以对单个smorf进行翻译评分。鉴定翻译的smorf是研究微蛋白功能的第一步。有关该协议的使用和执行的完整细节,请参阅马丁内斯等人。高质量Ribo-seq文库的准备对测序的Ribo-seq数据进行处理、比对和质量检查从转录组中创建ORF数据库用于翻译预测使用RibORF软件包对小ORF翻译进行评分出版商注:进行任何实验方案都需要遵守当地机构的实验室安全和伦理指导方针。小开放阅读框(smORF)编码的微蛋白是一类含有少于100-150个氨基酸的新型功能性生物分子。在这里,我们提出了一种在哺乳动物系统全基因组范围内鉴定翻译smorf的方案。我们描述了生成核糖体分析(Ribo-seq)数据的步骤,转录组组装的计算机翻译以创建ORF数据库,以及Ribo-seq的计算分析以对单个smorf进行翻译评分。鉴定翻译的smorf是研究微蛋白功能的第一步。
Small open reading frame (smORF)-encoded microproteins, proteins containing less than 100–150 amino acids, are an emerging class of functional biomolecules. Here, we present a protocol for identifying translated smORFs in mammalian systems genome wide. We describe steps for generation of ribosome profiling (Ribo-seq) data, in silico translation of a transcriptome assembly to create an ORF database, and computational analysis of Ribo-seq to score individual smORFs for translation. Identification of translated smORFs is the first step to studying the functions of microproteins. For complete details on the use and execution of this protocol, please refer to Martinez et al. Preparation of high-quality Ribo-seq libraries Processing, aligning, and quality checks for sequenced Ribo-seq data Creation of ORF database from a transcriptome to use in translation prediction Scoring of small ORF translation using RibORF software package Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Small open reading frame (smORF)-encoded microproteins, proteins containing less than 100–150 amino acids, are an emerging class of functional biomolecules. Here, we present a protocol for identifying translated smORFs in mammalian systems genome wide. We describe steps for generation of ribosome profiling (Ribo-seq) data, in silico translation of a transcriptome assembly to create an ORF database, and computational analysis of Ribo-seq to score individual smORFs for translation. Identification of translated smORFs is the first step to studying the functions of microproteins.
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