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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Online Access: | https://hdl.handle.net/11250/3111684 https://doi.org/10.1007/s11248-023-00368-4 |
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ftimr:oai:imr.brage.unit.no:11250/3111684 2024-02-11T10:02:03+01:00 In vivo CRISPR/LbCas12a-mediated knock-in and knock-out in Atlantic salmon (Salmo salar L.) Raudstein, Mari Torsdotter Kjærner-Semb, Erik Nordtorp Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik 2023 application/pdf https://hdl.handle.net/11250/3111684 https://doi.org/10.1007/s11248-023-00368-4 eng eng Transgenic research. 2023, . urn:issn:0962-8819 https://hdl.handle.net/11250/3111684 https://doi.org/10.1007/s11248-023-00368-4 cristin:2192497 9 Transgenic research Peer reviewed Journal article 2023 ftimr https://doi.org/10.1007/s11248-023-00368-4 2024-01-17T23:47:42Z 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. publishedVersion Article in Journal/Newspaper Atlantic salmon Salmo salar Institute for Marine Research: Brage IMR Transgenic Research 32 6 513 521 |
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Institute for Marine Research: Brage IMR |
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English |
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. publishedVersion |
format |
Article in Journal/Newspaper |
author |
Raudstein, Mari Torsdotter Kjærner-Semb, Erik Nordtorp Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik |
spellingShingle |
Raudstein, Mari Torsdotter Kjærner-Semb, Erik Nordtorp 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.) |
author_facet |
Raudstein, Mari Torsdotter Kjærner-Semb, Erik Nordtorp Barvik, Morten Broll, Silje Straume, Anne Hege Edvardsen, Rolf Brudvik |
author_sort |
Raudstein, Mari Torsdotter |
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.) |
publishDate |
2023 |
url |
https://hdl.handle.net/11250/3111684 https://doi.org/10.1007/s11248-023-00368-4 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
9 Transgenic research |
op_relation |
Transgenic research. 2023, . urn:issn:0962-8819 https://hdl.handle.net/11250/3111684 https://doi.org/10.1007/s11248-023-00368-4 cristin:2192497 |
op_doi |
https://doi.org/10.1007/s11248-023-00368-4 |
container_title |
Transgenic Research |
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32 |
container_issue |
6 |
container_start_page |
513 |
op_container_end_page |
521 |
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1790597955095363584 |