单只小鼠卵母细胞的转录组分析

Transcriptome Profiling of Single Mouse Oocytes.

作者信息

Borensztein Maud, Syx Laurène, Servant Nicolas, Heard Edith

机构信息

Institut Curie, PSL Research University, CNRS UMR3215, INSERM U934, UPMC Paris-Sorbonne, Paris, 75005, France.

Institut Curie, PSL Research University, Mines Paris Tech, INSERM U900, Paris, 75005, France.

出版信息

Methods Mol Biol. 2018;1818:51-65. doi: 10.1007/978-1-4939-8603-3_7.

DOI:10.1007/978-1-4939-8603-3_7

PMID:29961255

Abstract

Single-cell RNA-sequencing (scRNAseq) enables the detection and quantification of mature RNAs in an individual cell. Assessing single cell transcriptomes can circumvent the limited amount of starting material obtained in oocytes or embryos, in particular when working with mutant mice. Here we outline our scRNAseq protocol to study mouse oocyte transcriptomes, derived from Tang et al., Nat Methods 6(5):377-382, 2009 . The method describes the different steps from single cell isolation and cDNA amplification to high-throughput sequencing. The bioinformatics pipeline used to analyze and compare genome-wide gene expression between individual oocytes is then described.

摘要

单细胞RNA测序（scRNAseq）能够检测和定量单个细胞中的成熟RNA。评估单细胞转录组可以规避从卵母细胞或胚胎中获得的起始材料数量有限的问题，特别是在研究突变小鼠时。在此，我们概述了我们用于研究小鼠卵母细胞转录组的scRNAseq方案，该方案源自Tang等人发表于《自然方法》2009年第6卷第5期第377 - 382页的文章。该方法描述了从单细胞分离、cDNA扩增到高通量测序的不同步骤。随后介绍了用于分析和比较单个卵母细胞之间全基因组基因表达的生物信息学流程。