Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes
Abstract Infectious and parasitic diseases have major negative economic and animal welfare impacts on aquaculture of salmonid species. Improved knowledge of the functional basis of host response and genetic resistance to these diseases is key to developing preventative and treatment options. Cell li...
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Springer Science and Business Media LLC
2020
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Online Access: | http://dx.doi.org/10.1007/s10126-020-09995-y https://link.springer.com/content/pdf/10.1007/s10126-020-09995-y.pdf https://link.springer.com/article/10.1007/s10126-020-09995-y/fulltext.html |
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crspringernat:10.1007/s10126-020-09995-y 2023-05-15T15:31:42+02:00 Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. Biotechnology and Biological Sciences Research Council Hendrix Genetics 2020 http://dx.doi.org/10.1007/s10126-020-09995-y https://link.springer.com/content/pdf/10.1007/s10126-020-09995-y.pdf https://link.springer.com/article/10.1007/s10126-020-09995-y/fulltext.html en eng Springer Science and Business Media LLC https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 CC-BY Marine Biotechnology volume 22, issue 5, page 717-724 ISSN 1436-2228 1436-2236 Applied Microbiology and Biotechnology journal-article 2020 crspringernat https://doi.org/10.1007/s10126-020-09995-y 2022-01-04T16:08:02Z Abstract Infectious and parasitic diseases have major negative economic and animal welfare impacts on aquaculture of salmonid species. Improved knowledge of the functional basis of host response and genetic resistance to these diseases is key to developing preventative and treatment options. Cell lines provide valuable models to study infectious diseases in salmonids, and genome editing using CRISPR/Cas systems provides an exciting avenue to evaluate the function of specific genes in those systems. While CRISPR/Cas editing has been successfully performed in a Chinook salmon cell line (CHSE-214), there are no reports to date of editing of cell lines derived from the most commercially relevant salmonid species Atlantic salmon and rainbow trout, which are difficult to transduce and therefore edit using lentivirus-mediated methods. In the current study, a method of genome editing of salmonid cell lines using ribonucleoprotein (RNP) complexes was optimised and tested in the most commonly used salmonid fish cell lines: Atlantic salmon (SHK-1 and ASK cell lines), rainbow trout (RTG-2) and Chinook salmon (CHSE-214). Electroporation of RNP based on either Cas9 or Cas12a was efficient at targeted editing of all the tested lines (typically > 90% cells edited), and the choice of enzyme expands the number of potential target sites for editing within the genomes of these species. These optimised protocols will facilitate functional genetic studies in salmonid cell lines, which are widely used as model systems for infectious diseases in aquaculture. Article in Journal/Newspaper Atlantic salmon Springer Nature (via Crossref) Marine Biotechnology 22 5 717 724 |
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Springer Nature (via Crossref) |
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English |
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Applied Microbiology and Biotechnology |
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Applied Microbiology and Biotechnology Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
topic_facet |
Applied Microbiology and Biotechnology |
description |
Abstract Infectious and parasitic diseases have major negative economic and animal welfare impacts on aquaculture of salmonid species. Improved knowledge of the functional basis of host response and genetic resistance to these diseases is key to developing preventative and treatment options. Cell lines provide valuable models to study infectious diseases in salmonids, and genome editing using CRISPR/Cas systems provides an exciting avenue to evaluate the function of specific genes in those systems. While CRISPR/Cas editing has been successfully performed in a Chinook salmon cell line (CHSE-214), there are no reports to date of editing of cell lines derived from the most commercially relevant salmonid species Atlantic salmon and rainbow trout, which are difficult to transduce and therefore edit using lentivirus-mediated methods. In the current study, a method of genome editing of salmonid cell lines using ribonucleoprotein (RNP) complexes was optimised and tested in the most commonly used salmonid fish cell lines: Atlantic salmon (SHK-1 and ASK cell lines), rainbow trout (RTG-2) and Chinook salmon (CHSE-214). Electroporation of RNP based on either Cas9 or Cas12a was efficient at targeted editing of all the tested lines (typically > 90% cells edited), and the choice of enzyme expands the number of potential target sites for editing within the genomes of these species. These optimised protocols will facilitate functional genetic studies in salmonid cell lines, which are widely used as model systems for infectious diseases in aquaculture. |
author2 |
Biotechnology and Biological Sciences Research Council Hendrix Genetics |
format |
Article in Journal/Newspaper |
author |
Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. |
author_facet |
Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. |
author_sort |
Gratacap, Remi L. |
title |
Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
title_short |
Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
title_full |
Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
title_fullStr |
Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
title_full_unstemmed |
Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
title_sort |
efficient genome editing in multiple salmonid cell lines using ribonucleoprotein complexes |
publisher |
Springer Science and Business Media LLC |
publishDate |
2020 |
url |
http://dx.doi.org/10.1007/s10126-020-09995-y https://link.springer.com/content/pdf/10.1007/s10126-020-09995-y.pdf https://link.springer.com/article/10.1007/s10126-020-09995-y/fulltext.html |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Marine Biotechnology volume 22, issue 5, page 717-724 ISSN 1436-2228 1436-2236 |
op_rights |
https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1007/s10126-020-09995-y |
container_title |
Marine Biotechnology |
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22 |
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5 |
container_start_page |
717 |
op_container_end_page |
724 |
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1766362221225443328 |