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

Remi Gratacap 1, * Tim Regan 1 Carola Dehler 2 Samuel Martin 2 Pierre Boudinot 3 Bertrand Collet 3 Ross D. Houston 1, *
* Corresponding author
1 The Roslin Institute
2 University of Aberdeen
3 VIM (UR 0892) - Virologie et Immunologie Moléculaires
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.
Keywords : CRISPR Lentivirus Gene editing CHSE Salmon Disease resistance
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Journal articles
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https://hal.inrae.fr/hal-03129758
Contributor : Bertrand Collet Connect in order to contact the contributor
Submitted on : Tuesday, December 14, 2021 - 11:21:27 AM
Last modification on : Tuesday, April 5, 2022 - 11:38:02 AM
Long-term archiving on: : Tuesday, March 15, 2022 - 6:42:59 PM

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UVSQ | INRAE | VIM | GS-BIOSPHERA | GS-LIFE-SCIENCES-HEALTH | GS-HEALTH-DRUG-SCIENCES | UNIV-PARIS-SACLAY

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Remi Gratacap, Tim Regan, Carola Dehler, Samuel Martin, Pierre Boudinot, et al.. Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system. BMC Biotechnology, BioMed Central, 2020, 20 (1), ⟨10.1186/s12896-020-00626-x⟩. ⟨hal-03129758⟩

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