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

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

Research output: Contribution to journalArticlepeer-review

Abstract

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
JournalBMC Biotechnology
Early online date20 Jun 2020
DOIs
Publication statusE-pub ahead of print - 20 Jun 2020

Keywords

  • CRISPR
  • Lentivirus
  • Gene editing
  • CHSE
  • Salmon
  • Disease resistance

Fingerprint

Dive into the research topics of 'Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system'. Together they form a unique fingerprint.

Cite this