Molecular aspects of zinc finger nucleases (ZFNs)-mediated gene editing in rat embryos

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

This chapter contains a collection of protocols involved in using ZFNs to create rat models with various types of genome editing, including simple knockout, point mutation, large deletions, floxing, and insertions. The protocols cover ZFN and donor design criteria, in vitro transcription of ZFNs, validation of ZFNs activity in cultured cells, RNA stability test, microinjection sample preparation, genotyping and in vitro confirmation of floxed alleles, and Southern blot analysis, most of which are not limited to using ZFNs. Instead they apply to model creation in general. When appropriate, a comparison between ZFNs and CRISPR is provided. Standard pronuclear microinjection per se is not discussed.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages295-312
Number of pages18
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1874
ISSN (Print)1064-3745

Keywords

  • Conditional knockout
  • Homology directed repair
  • Knockin
  • Knockout
  • Nonhomologous end joining
  • Pronuclear microinjection
  • Single-cell embryos
  • Zinc finger nucleases

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