Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes
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 provi...
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ftpubmed:oai:pubmedcentral.nih.gov:7520412 2023-05-15T15:31:46+02:00 Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. 2020-09-18 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520412/ http://www.ncbi.nlm.nih.gov/pubmed/32946000 https://doi.org/10.1007/s10126-020-09995-y en eng Springer US http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520412/ http://www.ncbi.nlm.nih.gov/pubmed/32946000 http://dx.doi.org/10.1007/s10126-020-09995-y © 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 Commons licence, 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. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. CC-BY Mar Biotechnol (NY) Original Article Text 2020 ftpubmed https://doi.org/10.1007/s10126-020-09995-y 2020-10-18T00:19:48Z 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10126-020-09995-y) contains supplementary material, which is available to authorized users. Text Atlantic salmon PubMed Central (PMC) Marine Biotechnology 22 5 717 724 |
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Original Article Gratacap, Remi L. Jin, Ye Hwa Mantsopoulou, Marina Houston, Ross D. Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes |
topic_facet |
Original Article |
description |
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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10126-020-09995-y) contains supplementary material, which is available to authorized users. |
format |
Text |
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 US |
publishDate |
2020 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520412/ http://www.ncbi.nlm.nih.gov/pubmed/32946000 https://doi.org/10.1007/s10126-020-09995-y |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Mar Biotechnol (NY) |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520412/ http://www.ncbi.nlm.nih.gov/pubmed/32946000 http://dx.doi.org/10.1007/s10126-020-09995-y |
op_rights |
© 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 Commons licence, 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. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1007/s10126-020-09995-y |
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Marine Biotechnology |
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22 |
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5 |
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717 |
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724 |
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