Targeted gene knock out using nuclease-assisted vector integration: Hemi- and Homozygous deletion of JAG1

Michael Gapinske, Nathan Tague, Jackson Winter, Gregory H. Underhill, Pablo Perez-Pinera

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Gene editing technologies are revolutionizing fields such as biomedicine and biotechnology by providing a simple means to manipulate the genetic makeup of essentially any organism. Gene editing tools function by introducing double-stranded breaks at targeted sites within the genome, which the host cells repair preferentially by Non-Homologous End Joining. While the technologies to introduce double-stranded breaks have been extensively optimized, this progress has not been matched by the development of methods to integrate heterologous DNA at the target sites or techniques to detect and isolate cells that harbor the desired modification. We present here a technique for rapid introduction of vectors at target sites in the genome that enables efficient isolation of successfully edited cells.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages233-248
Number of pages16
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1772
ISSN (Print)1064-3745

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Keywords

  • CRISPR-Cas9
  • Gene editing
  • Gene knock out
  • Genome engineering

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Gapinske, M., Tague, N., Winter, J., Underhill, G. H., & Perez-Pinera, P. (2018). Targeted gene knock out using nuclease-assisted vector integration: Hemi- and Homozygous deletion of JAG1. In Methods in Molecular Biology (pp. 233-248). (Methods in Molecular Biology; Vol. 1772). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7795-6_13