Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System

Jinchao Gu, Huseyin Sumer, Brett Cromer

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

3 Citations (Scopus)

Abstract

The piggyBac transposon system has been adapted to be a highly efficient genome engineering tool for transgenesis of eukaryotic cells and organisms. As with other methods of transgenesis, incorporation of an inducible promoter, such as a tetracycline-responsive element, enables inducible transgene expression. Here, we describe an efficient method of using the piggyBac system to create stably transfected mammalian cell lines, including inducible transgene expression. Gibson assembly is used to construct the required vectors as it enables multiple DNA fragments to be seamlessly assembled in a single isothermal reaction. We demonstrate an application of this approach to generate a stably transfected pluripotent stem cell line that can be induced to express a transcription factor transgene and rapidly differentiate into neurons in a single step.

Original languageEnglish
Title of host publicationApplications of Genome Modulation and Editing
EditorsPaul John Verma, Huseyin Sumer, Jun Liu
Place of PublicationUSA
PublisherHumana Press
Chapter3
Pages49-66
Number of pages18
ISBN (Electronic)9781071623015
ISBN (Print)9781071623008
DOIs
Publication statusPublished - 2022
Externally publishedYes

Publication series

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

Keywords

  • Gibson assembly
  • Inducible transgene expression
  • Neuronal differentiation
  • PiggyBac
  • Stem cells

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