Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system

Remi L Gratacap, Tim Regan, Carola E Dehler, Samuel A M Martin, Pierre Boudinot, Bertrand Collet, Ross D Houston

Research output: Contribution to journalArticle

2 Citations (Scopus)
1 Downloads (Pure)

Abstract

BACKGROUND: Genome editing is transforming bioscience research, but its application to non-model organisms, such as farmed animal species, requires optimisation. Salmonids are the most important aquaculture species by value, and improving genetic resistance to infectious disease is a major goal. However, use of genome editing to evaluate putative disease resistance genes in cell lines, and the use of genome-wide CRISPR screens is currently limited by a lack of available tools and techniques.

RESULTS: In the current study, we developed an optimised protocol using lentivirus transduction for efficient integration of constructs into the genome of a Chinook salmon (Oncorhynchus tshwaytcha) cell line (CHSE-214). As proof-of-principle, two target genes were edited with high efficiency in an EGFP-Cas9 stable CHSE cell line; specifically, the exogenous, integrated EGFP and the endogenous RIG-I locus. Finally, the effective use of antibiotic selection to enrich the successfully edited targeted population was demonstrated.

CONCLUSIONS: The optimised lentiviral-mediated CRISPR method reported here increases possibilities for efficient genome editing in salmonid cells, in particular for future applications of genome-wide CRISPR screens for disease resistance.

Original languageEnglish
Article number35
Number of pages9
JournalBMC Biotechnology
Volume20
DOIs
Publication statusPublished - 23 Jun 2020

Keywords

  • CRISPR
  • lentivirus
  • gene editing
  • CHSE
  • salmon
  • disease resistance
  • Disease resistance
  • Salmon
  • Gene editing
  • Lentivirus
  • VIRUS
  • TRANSDUCTION
  • FUTURE
  • GENES
  • VECTORS

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