Targeting the AAVS1 Site by CRISPR/Cas9 with an Inducible Transgene Cassette for the Neuronal Differentiation of Human Pluripotent Stem Cells

Jinchao Gu, Ben Rollo, Huseyin Sumer, Brett Cromer

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

1 Citation (Scopus)

Abstract

CRISPR/Cas9 system is a powerful genome-editing technology for studying genetics and cell biology. Safe harbor sites are ideal genomic locations for transgene integration with minimal interference in cellular functions. Gene targeting of the AAVS1 locus enables stable transgene expression without phenotypic effects in host cells. Here, we describe the strategy for targeting the AAVS1 site with an inducible Neurogenin-2 (Ngn2) donor template by CRISPR/Cas9 in hiPSCs, which facilitates generation of an inducible cell line that can rapidly and homogenously differentiate into excitatory neurons.

Original languageEnglish
Title of host publicationApplications of Genome Modulation and Editing
EditorsPaul John Verma, Huseyin Sumer, Jun Liu
Place of PublicationNew York USA
PublisherHumana Press
Chapter6
Pages99-114
Number of pages16
ISBN (Electronic)9781071623015
ISBN (Print)9781071623008
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2495
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • AAVS1
  • CRISPR/Cas9
  • Neuronal differentiation
  • Ngn2
  • Stem cells

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