Unknown

Dataset Information

0

Cytoplasmic Injection of Zygotes to Genome Edit Naturally Occurring Sequence Variants Into Bovine Embryos.


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.

SUBMITTER: Wei J 

PROVIDER: S-EPMC9310181 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC5354444 | biostudies-literature
| S-EPMC11310320 | biostudies-literature
| S-EPMC2716050 | biostudies-literature
| S-EPMC6639132 | biostudies-literature
| S-EPMC10237670 | biostudies-literature
2017-09-01 | GSE73276 | GEO
| S-EPMC5705543 | biostudies-literature
| S-EPMC1540737 | biostudies-literature
| S-EPMC3503569 | biostudies-literature
| S-EPMC4617447 | biostudies-literature