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

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    Rights statement: © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commonslicence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder ...

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

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.

    Research areas

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

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