In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.)
Genome editing using the CRISPR/Cas system offers the potential to enhance current breeding programs and introduce desirable genetic traits, including disease resistance, in salmon aquaculture. Several nucleases are available using this system, displaying differences regarding structure, cleavage, a...
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ftpubmed:oai:pubmedcentral.nih.gov:10713776 2024-01-14T10:05:25+01:00 In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) Raudstein, Mari Kjærner-Semb, Erik Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik 2023-09-21 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10713776/ http://www.ncbi.nlm.nih.gov/pubmed/37733197 https://doi.org/10.1007/s11248-023-00368-4 en eng Springer International Publishing http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10713776/ http://www.ncbi.nlm.nih.gov/pubmed/37733197 http://dx.doi.org/10.1007/s11248-023-00368-4 © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . Transgenic Res Research Text 2023 ftpubmed https://doi.org/10.1007/s11248-023-00368-4 2023-12-17T01:54:36Z Genome editing using the CRISPR/Cas system offers the potential to enhance current breeding programs and introduce desirable genetic traits, including disease resistance, in salmon aquaculture. Several nucleases are available using this system, displaying differences regarding structure, cleavage, and PAM requirement. Cas9 is well established in Atlantic salmon, but Cas12a has yet to be tested in vivo in this species. In the present work, we microinjected salmon embryos with LbCas12a ribonucleoprotein complexes targeting the pigmentation gene solute carrier family 45 member 2 (slc45a2). Using CRISPR/LbCas12a, we were able to knock-out slc45a2 and knock-in a FLAG sequence element by providing single-stranded DNA templates. High-throughput sequencing revealed perfect HDR rates up to 34.3% and 54.9% in individual larvae using either target or non-target strand template design, respectively. In this work, we demonstrate the in vivo application of CRISPR/LbCas12a in Atlantic salmon, expanding the toolbox for editing the genome of this important aquaculture species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11248-023-00368-4. Text Atlantic salmon Salmo salar PubMed Central (PMC) Transgenic Research 32 6 513 521 |
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Research Raudstein, Mari Kjærner-Semb, Erik Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
topic_facet |
Research |
description |
Genome editing using the CRISPR/Cas system offers the potential to enhance current breeding programs and introduce desirable genetic traits, including disease resistance, in salmon aquaculture. Several nucleases are available using this system, displaying differences regarding structure, cleavage, and PAM requirement. Cas9 is well established in Atlantic salmon, but Cas12a has yet to be tested in vivo in this species. In the present work, we microinjected salmon embryos with LbCas12a ribonucleoprotein complexes targeting the pigmentation gene solute carrier family 45 member 2 (slc45a2). Using CRISPR/LbCas12a, we were able to knock-out slc45a2 and knock-in a FLAG sequence element by providing single-stranded DNA templates. High-throughput sequencing revealed perfect HDR rates up to 34.3% and 54.9% in individual larvae using either target or non-target strand template design, respectively. In this work, we demonstrate the in vivo application of CRISPR/LbCas12a in Atlantic salmon, expanding the toolbox for editing the genome of this important aquaculture species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11248-023-00368-4. |
format |
Text |
author |
Raudstein, Mari Kjærner-Semb, Erik Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik |
author_facet |
Raudstein, Mari Kjærner-Semb, Erik Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik |
author_sort |
Raudstein, Mari |
title |
In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
title_short |
In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
title_full |
In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
title_fullStr |
In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
title_full_unstemmed |
In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) |
title_sort |
in vivo crispr/lbcas12a-mediated knock-in and knock-out in atlantic salmon (salmo salar l.) |
publisher |
Springer International Publishing |
publishDate |
2023 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10713776/ http://www.ncbi.nlm.nih.gov/pubmed/37733197 https://doi.org/10.1007/s11248-023-00368-4 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
Transgenic Res |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10713776/ http://www.ncbi.nlm.nih.gov/pubmed/37733197 http://dx.doi.org/10.1007/s11248-023-00368-4 |
op_rights |
© The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . |
op_doi |
https://doi.org/10.1007/s11248-023-00368-4 |
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Transgenic Research |
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32 |
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6 |
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513 |
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521 |
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1788059781326438400 |