.png){kind=icon}
返回
Applying thiouracil tagging to mouse transcriptome analysis
.png){kind=icon}
OA
AI
DOI：10.1038/nprot.2014.023.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Transcriptional profiling is a powerful approach for studying mouse development, physiology and disease models. Here we describe a protocol for mouse thiouracil tagging (TU tagging), a transcriptome analysis technology that includes in vivo covalent labeling, purification and analysis of cell type-specific RNA. TU tagging enables the isolation of RNA from a given cell population of a complex tissue, avoiding transcriptional changes induced by cell isolation trauma, as well as the identification of actively transcribed RNAs and not preexisting transcripts. Therefore, in contrast to other cell-specific transcriptional profiling methods based on the purification of tagged ribosomes or nuclei, TU tagging provides a direct examination of transcriptional regulation. We describe how to (i) deliver 4-thiouracil to transgenic mice to thio-label cell lineage-specific transcripts, (ii) purify TU-tagged RNA and prepare libraries for Illumina sequencing and (iii) follow a straightforward bioinformatics workflow to identify cell type-enriched or differentially expressed genes. Tissue containing TU-tagged RNA can be obtained in 1 d, RNA-seq libraries can be generated within 2 d and, after sequencing, an initial bioinformatics analysis can be completed in 1 additional day.
Keyword:
TRANSLATIONAL PROFILING APPROACH
MICROARRAY ANALYSIS
MESSENGER-RNA
EXPRESSION
期刊
IF:
16
论文数:
4.0K
被引数:
5.6W
机构
引用论文
分享收藏Comparative analysis of RNA sequencing methods for degraded or low-input samples.png)
NATURE METHODS
OAAI
NATURE METHODSIF32.1
OAAI分享收藏Cell-type-specific isolation of ribosome-associated mRNA from complex tissues来自复杂组织的核糖体相关mRNA的细胞类型特异性隔离OAAI
OAAI分享收藏Biosynthetic labeling of RNA with uracil phosphoribosyltransferase allows cell-specific microarray analysis of mRNA synthesis and decay用尿嘧啶磷酸核糖基转移酶对RNA进行生物合成标记，可以对mRNA的合成和衰变进行细胞特异性微阵列分析。NATURE BIOTECHNOLOGY
NATURE BIOTECHNOLOGYIF41.7
PREAI
分享收藏
查看更多内容