Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein (RNP) electroporation.

ABSTRACT:

Background

Genome editing has been considered as powerful tool in agricultural fields. However, genome editing progress in cattle has not been fast as in other mammal species, for some disadvantages including long gestational periods, single pregnancy, and high raising cost. Furthermore, technically demanding methods such as microinjection and somatic cell nuclear transfer (SCNT) are needed for gene editing in cattle. In this point of view, electroporation in embryos has been risen as an alternative.

Results

First, editing efficiency of our electroporation methods were tested for embryos. Presence of mutation on embryo was confirmed by T7E1 assay. With first combination, mutation rates for MSTN and PRNP were 57.6% ± 13.7% and 54.6% ± 13.5%, respectively. In case of MSTN/BLG,

SUBMITTER: Gim GM 

PROVIDER: S-EPMC10404370 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature