Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors
Abstract Background New breeding technologies (NBT) using CRISPR/Cas9-induced homology directed repair (HDR) has the potential to expedite genetic improvement in aquaculture. The long generation time in Atlantic salmon makes breeding an unattractive solution to obtain homozygous mutants and improvin...
Published in: | BMC Genomics |
---|---|
Main Authors: | , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
BMC
2021
|
Subjects: | |
Online Access: | https://doi.org/10.1186/s12864-021-07823-8 https://doaj.org/article/de4d172cfe394feca90b2722376b76e6 |
id |
ftdoajarticles:oai:doaj.org/article:de4d172cfe394feca90b2722376b76e6 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:de4d172cfe394feca90b2722376b76e6 2023-05-15T15:31:42+02:00 Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors Anne Hege Straume Erik Kjærner-Semb Kai Ove Skaftnesmo Hilal Güralp Simon Lillico Anna Wargelius Rolf Brudvik Edvardsen 2021-07-01T00:00:00Z https://doi.org/10.1186/s12864-021-07823-8 https://doaj.org/article/de4d172cfe394feca90b2722376b76e6 EN eng BMC https://doi.org/10.1186/s12864-021-07823-8 https://doaj.org/toc/1471-2164 doi:10.1186/s12864-021-07823-8 1471-2164 https://doaj.org/article/de4d172cfe394feca90b2722376b76e6 BMC Genomics, Vol 22, Iss 1, Pp 1-8 (2021) HDR ODN Gene editing Knock-in Aquaculture New breeding technologies Biotechnology TP248.13-248.65 Genetics QH426-470 article 2021 ftdoajarticles https://doi.org/10.1186/s12864-021-07823-8 2022-12-31T13:57:50Z Abstract Background New breeding technologies (NBT) using CRISPR/Cas9-induced homology directed repair (HDR) has the potential to expedite genetic improvement in aquaculture. The long generation time in Atlantic salmon makes breeding an unattractive solution to obtain homozygous mutants and improving the rates of perfect HDR in founder (F0) fish is thus required. Genome editing can represent small DNA changes down to single nucleotide replacements (SNR). This enables edits such as premature stop codons or single amino acid changes and may be used to obtain fish with traits favorable to aquaculture, e.g. disease resistance. A method for SNR has not yet been demonstrated in salmon. Results Using CRISPR/Cas9 and asymmetrical ODNs, we were able to perform precise SNR and introduce a premature stop codon in dnd in F0 salmon. Deep sequencing demonstrated up to 59.2% efficiency in single embryos. In addition, using the same asymmetrical ODN design, we inserted a FLAG element into slc45a2 and dnd, showing high individual perfect HDR efficiencies (up to 36.7 and 32.7%, respectively). Conclusions In this work, we demonstrate that precise SNR and knock-in (KI) can be performed in F0 salmon embryos using asymmetrical oligonucleotide (ODN) donors. We suggest that HDR-induced SNR can be applied as a powerful NBT, allowing efficient introgression of favorable alleles and bypassing challenges associated with traditional selective breeding. Article in Journal/Newspaper Atlantic salmon Directory of Open Access Journals: DOAJ Articles BMC Genomics 22 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
HDR ODN Gene editing Knock-in Aquaculture New breeding technologies Biotechnology TP248.13-248.65 Genetics QH426-470 |
spellingShingle |
HDR ODN Gene editing Knock-in Aquaculture New breeding technologies Biotechnology TP248.13-248.65 Genetics QH426-470 Anne Hege Straume Erik Kjærner-Semb Kai Ove Skaftnesmo Hilal Güralp Simon Lillico Anna Wargelius Rolf Brudvik Edvardsen Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
topic_facet |
HDR ODN Gene editing Knock-in Aquaculture New breeding technologies Biotechnology TP248.13-248.65 Genetics QH426-470 |
description |
Abstract Background New breeding technologies (NBT) using CRISPR/Cas9-induced homology directed repair (HDR) has the potential to expedite genetic improvement in aquaculture. The long generation time in Atlantic salmon makes breeding an unattractive solution to obtain homozygous mutants and improving the rates of perfect HDR in founder (F0) fish is thus required. Genome editing can represent small DNA changes down to single nucleotide replacements (SNR). This enables edits such as premature stop codons or single amino acid changes and may be used to obtain fish with traits favorable to aquaculture, e.g. disease resistance. A method for SNR has not yet been demonstrated in salmon. Results Using CRISPR/Cas9 and asymmetrical ODNs, we were able to perform precise SNR and introduce a premature stop codon in dnd in F0 salmon. Deep sequencing demonstrated up to 59.2% efficiency in single embryos. In addition, using the same asymmetrical ODN design, we inserted a FLAG element into slc45a2 and dnd, showing high individual perfect HDR efficiencies (up to 36.7 and 32.7%, respectively). Conclusions In this work, we demonstrate that precise SNR and knock-in (KI) can be performed in F0 salmon embryos using asymmetrical oligonucleotide (ODN) donors. We suggest that HDR-induced SNR can be applied as a powerful NBT, allowing efficient introgression of favorable alleles and bypassing challenges associated with traditional selective breeding. |
format |
Article in Journal/Newspaper |
author |
Anne Hege Straume Erik Kjærner-Semb Kai Ove Skaftnesmo Hilal Güralp Simon Lillico Anna Wargelius Rolf Brudvik Edvardsen |
author_facet |
Anne Hege Straume Erik Kjærner-Semb Kai Ove Skaftnesmo Hilal Güralp Simon Lillico Anna Wargelius Rolf Brudvik Edvardsen |
author_sort |
Anne Hege Straume |
title |
Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
title_short |
Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
title_full |
Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
title_fullStr |
Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
title_full_unstemmed |
Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors |
title_sort |
single nucleotide replacement in the atlantic salmon genome using crispr/cas9 and asymmetrical oligonucleotide donors |
publisher |
BMC |
publishDate |
2021 |
url |
https://doi.org/10.1186/s12864-021-07823-8 https://doaj.org/article/de4d172cfe394feca90b2722376b76e6 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
BMC Genomics, Vol 22, Iss 1, Pp 1-8 (2021) |
op_relation |
https://doi.org/10.1186/s12864-021-07823-8 https://doaj.org/toc/1471-2164 doi:10.1186/s12864-021-07823-8 1471-2164 https://doaj.org/article/de4d172cfe394feca90b2722376b76e6 |
op_doi |
https://doi.org/10.1186/s12864-021-07823-8 |
container_title |
BMC Genomics |
container_volume |
22 |
container_issue |
1 |
_version_ |
1766362218392190976 |