Cell-specific RNA purification to study translatomes of mouse central nervous system.
Cell-specific RNA purification to study translatomes of mouse central nervous system.
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利用细胞特异性RNA纯化技术研究小鼠中枢神经系统的翻译体。
DOI：
10.1016/j.xpro.2022.101397
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发表时间：
2022-06-17
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细胞特异性RNA测序已经彻底改变了细胞生物学的研究。在这里，我们提出了一个协议，以评估细胞特异性translatomes的遗传靶向细胞类型。我们专注于星形胶质细胞和描述RNA纯化使用RiboTag工具。与单细胞RNA测序不同，这种方法允许高测序深度来检测低表达基因，并探索在亚细胞区室中翻译的RNA。此外，它避免了对来自易受细胞分离程序影响的细胞的转录物的低估。该方案可应用于多种细胞类型。有关本方案使用和执行的完整详细信息，请参见、和。研究小鼠脑细胞中细胞特异性翻译体的方案使用RiboTag工具从遗传靶向细胞中纯化RNA推荐抗体和寡核苷酸以评估脑细胞特异性用于生成最可靠数据的样品质量评估提示细胞特异性RNA测序彻底改变了细胞生物学研究。在这里，我们提出了一个协议，以评估细胞特异性translatomes的遗传靶向细胞类型。我们专注于星形胶质细胞和描述RNA纯化使用RiboTag工具。与单细胞RNA测序不同，这种方法允许高测序深度来检测低表达基因，并探索在亚细胞区室中翻译的RNA。此外，它避免了对来自易受细胞分离程序影响的细胞的转录物的低估。该方案可应用于多种细胞类型。
Cell-specific RNA sequencing has revolutionized the study of cell biology. Here, we present a protocol to assess cell-specific translatomes of genetically targeted cell types. We focus on astrocytes and describe RNA purification using RiboTag tools. Unlike single-cell RNA sequencing, this approach allows high sequencing depth to detect low expression genes, and the exploration of RNAs translated in subcellular compartments. Furthermore, it avoids underestimation of transcripts from cells susceptible to cell isolation procedures. The protocol can be applied to a variety of cell types. For complete details on the use and execution of this protocol, please refer to, and. A protocol for investigating cell-specific translatomes in mouse brain cells Purification of RNA from genetically targeted cells using RiboTag tools Recommended antibodies and oligonucleotides to assess brain-cell specificity Sample quality assessment tips for generating the most reliable data Cell-specific RNA sequencing has revolutionized the study of cell biology. Here, we present a protocol to assess cell-specific translatomes of genetically targeted cell types. We focus on astrocytes and describe RNA purification using RiboTag tools. Unlike single-cell RNA sequencing, this approach allows high sequencing depth to detect low expression genes, and the exploration of RNAs translated in subcellular compartments. Furthermore, it avoids underestimation of transcripts from cells susceptible to cell isolation procedures. The protocol can be applied to a variety of cell types.