Dissecting Transcriptional Heterogeneity in Pluripotency: Single Cell Analysis of Mouse Embryonic Stem Cells View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2016-04-23

AUTHORS

Ana M. V. Guedes , Domingos Henrique , Elsa Abranches

ABSTRACT

Mouse Embryonic Stem cells (mESCs) show heterogeneous and dynamic expression of important pluripotency regulatory factors. Single-cell analysis has revealed the existence of cell-to-cell variability in the expression of individual genes in mESCs. Understanding how these heterogeneities are regulated and what their functional consequences are is crucial to obtain a more comprehensive view of the pluripotent state.In this chapter we describe how to analyze transcriptional heterogeneity by monitoring gene expression of Nanog, Oct4, and Sox2, using single-molecule RNA FISH in single mESCs grown in different cell culture medium. We describe in detail all the steps involved in the protocol, from RNA detection to image acquisition and processing, as well as exploratory data analysis. More... »

PAGES

101-119

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/7651_2016_356

DOI

http://dx.doi.org/10.1007/7651_2016_356

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1030195539

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27106496


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