Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system
| Autor: | Bertrand Collet, Pierre Boudinot, Carola E. Dehler, Ross D. Houston, Remi L. Gratacap, Samuel A.M. Martin, Tim Regan |
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| Přispěvatelé: | The Roslin Institute, University of Aberdeen, Virologie et Immunologie Moléculaires (VIM (UR 0892)), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Biotechnology and Biological Sciences Research Council (BB/R008612/1, BB/S004343/1 and RG, grant BB/R008973/1), Institute Strategic Programme Grants (BBS/E/D/20002172, BBS/E/D/30002275 and BBS/E/D/10002070) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Cell Survival
lcsh:Biotechnology CRISPR-Associated Proteins Population Locus (genetics) Computational biology Biology Gene editing Genome Cell Line 03 medical and health sciences Transduction (genetics) 0302 clinical medicine Genome editing Salmon lcsh:TP248.13-248.65 Animals CRISPR Clustered Regularly Interspaced Short Palindromic Repeats education Gene 030304 developmental biology 0303 health sciences education.field_of_study Disease resistance Methodology Article Lentivirus CHSE Infectious disease (medical specialty) 030220 oncology & carcinogenesis [SDV.IMM]Life Sciences [q-bio]/Immunology CRISPR-Cas Systems 030217 neurology & neurosurgery Salmonidae Biotechnology |
| Zdroj: | BMC Biotechnology BMC Biotechnology, BioMed Central, 2020, 20 (1), ⟨10.1186/s12896-020-00626-x⟩ Gratacap, R, Regan, T, Dehler, C E, Martin, S A M, Boudinot, P, Collet, B & Houston, R 2020, ' Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system ', BMC Biotechnology . https://doi.org/10.1186/s12896-020-00626-x BMC Biotechnology, Vol 20, Iss 1, Pp 1-9 (2020) |
| ISSN: | 1472-6750 |
| DOI: | 10.1186/s12896-020-00626-x⟩ |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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