dc.identifier.uri |
http://dx.doi.org/10.15488/13006 |
|
dc.identifier.uri |
https://www.repo.uni-hannover.de/handle/123456789/13110 |
|
dc.contributor.author |
Wei, Jingwei
|
|
dc.contributor.author |
Brophy, Brigid
|
|
dc.contributor.author |
Cole, Sally-Ann
|
|
dc.contributor.author |
Moormann, Jannis
|
|
dc.contributor.author |
Boch, Jens
|
|
dc.contributor.author |
Laible, Gӧtz
|
|
dc.date.accessioned |
2022-11-14T07:13:46Z |
|
dc.date.available |
2022-11-14T07:13:46Z |
|
dc.date.issued |
2022 |
|
dc.identifier.citation |
Wei, J.; Brophy, B.; Cole, S.-A.; Moormann, J.; Boch, J. et al.: Cytoplasmic Injection of Zygotes to Genome Edit Naturally Occurring Sequence Variants Into Bovine Embryos. In: Frontiers in genetics 13 (2022), 925913. DOI: https://doi.org/10.3389/fgene.2022.925913 |
|
dc.description.abstract |
Genome editing provides opportunities to improve current cattle breeding strategies through targeted introduction of natural sequence variants, accelerating genetic gain. This can be achieved by harnessing homology-directed repair mechanisms following editor-induced cleavage of the genome in the presence of a repair template. Introducing the genome editors into zygotes and editing in embryos has the advantage of uncompromised development into live animals and alignment with contemporary embryo-based improvement practices. In our study, we investigated the potential to introduce sequence variants, known from the pre-melanosomal protein 17 (PMEL) and prolactin receptor (PRLR) genes, and produce non-mosaic, edited embryos, completely converted into the precision genotype. Injection of gRNA/Cas9 editors into bovine zygotes to introduce a 3 bp deletion variant into the PMEL gene produced up to 11% fully converted embryos. The conversion rate was increased to up to 48% with the use of TALEN but only when delivered by plasmid. Testing three gRNA/Cas9 editors in the context of several known PRLR sequence variants, different repair template designs and delivery as DNA, RNA or ribonucleoprotein achieved full conversion rates up to 8%. Furthermore, we developed a biopsy-based screening strategy for non-mosaic embryos which has the potential for exclusively producing non-mosaic animals with intended precision edits. Copyright © 2022 Wei, Brophy, Cole, Moormann, Boch and Laible. |
eng |
dc.language.iso |
eng |
|
dc.publisher |
Lausanne : Frontiers Media |
|
dc.relation.ispartofseries |
Frontiers in genetics 13 (2022) |
|
dc.rights |
CC BY 4.0 Unported |
|
dc.rights.uri |
https://creativecommons.org/licenses/by/4.0/ |
|
dc.subject |
Cas9 |
eng |
dc.subject |
cattle |
eng |
dc.subject |
genome editing |
eng |
dc.subject |
homology-directed repair |
eng |
dc.subject |
PMEL |
eng |
dc.subject |
PRLR |
eng |
dc.subject |
slick |
eng |
dc.subject |
TALEN |
eng |
dc.subject.ddc |
570 | Biowissenschaften, Biologie
|
ger |
dc.title |
Cytoplasmic Injection of Zygotes to Genome Edit Naturally Occurring Sequence Variants Into Bovine Embryos |
eng |
dc.type |
Article |
|
dc.type |
Text |
|
dc.relation.essn |
1664-8021 |
|
dc.relation.doi |
https://doi.org/10.3389/fgene.2022.925913 |
|
dc.bibliographicCitation.volume |
13 |
|
dc.bibliographicCitation.firstPage |
925913 |
|
dc.description.version |
publishedVersion |
|
tib.accessRights |
frei zug�nglich |
|