Gene Editing in Primary Cells of Cattle and Pig

Petra Vochozkova, Kilian Simmet, Eva Maria Jemiller, Annegret Wünsch, Nikolai Klymiuk

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

5 Scopus citations

Abstract

Gene Editing by CRISPR/Cas has revolutionized many aspects of biotechnology within a short period of time. This is also true for the genetic manipulation of livestock species, but their specific challenges such as the lack of stem cells, the limited proliferative capacity of primary cells, and the genetic diversity of the pig and cattle populations need consideration when CRISPR/Cas is applied. Here we present guidelines for CRISPRing primary cells in pig and cattle, with a specific focus on testing gRNA in vitro, on generating single cell clones, and on identifying modifications in single cell clones.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages271-289
Number of pages19
DOIs
StatePublished - 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1961
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CRISPR/Cas
  • Primary cells
  • Single cell clones
  • gRNA

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