Genetic tools for self-organizing culture of mouse embryonic stem cells via small regulatory RNA-mediated technologies, CRISPR/Cas9, and inducible RNAi

Nozomu Takata*, Eriko Sakakura, Tetsushi Sakuma, Takashi Yamamoto

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Approaches to investigate gene functions in experimental biology are becoming more diverse and reliable. Furthermore, several kinds of tissues and organs that possess their original identities can be generated in petri dishes from stem cells including embryonic, adult and induced pluripotent stem cells. Researchers now have several choices of experimental methods and their combinations to analyze gene functions in various biological systems. Here, as an example we describe one of the better protocols, which combines three-dimensional embryonic stem cell culture with small regulatory RNA-mediated technologies, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), and inducible RNA interference (RNAi). This protocol allows investigation of genes of interest to better understand gene functions in target tissues (or organs) during in vitro development.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages269-292
Number of pages24
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1622
ISSN (Print)1064-3745

Keywords

  • CRISPR/Cas9
  • Embryonic stem cell
  • Epiblast
  • Gene function
  • RNA interference
  • Small regulatory RNA
  • Three-dimensional culture

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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