Cytoplasmic injection of zygotes to genome edit naturally occurring sequence variants into bovine embryos
- verfasst von
- Jingwei Wei, Brigid Brophy, Sally Ann Cole, Jannis Moormann, Jens Boch, Gӧtz Laible
- 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.
- Organisationseinheit(en)
-
Abteilung Pflanzenmolekularbiologie und Pflanzenproteomik
Abteilung Pflanzenbiotechnologie
- Externe Organisation(en)
-
AgResearch
University of Auckland
- Typ
- Artikel
- Journal
- Frontiers in Genetics
- Band
- 13
- ISSN
- 1664-8021
- Publikationsdatum
- 11.07.2022
- Publikationsstatus
- Veröffentlicht
- Peer-reviewed
- Ja
- ASJC Scopus Sachgebiete
- Molekularmedizin, Genetik, Genetik (klinisch)
- Elektronische Version(en)
-
https://doi.org/10.3389/fgene.2022.925913 (Zugang:
Offen)