Optimization of CRISPR/Cas9 Gene Editing System in Sheep (<i>Ovis aries</i>) Oocytes via Microinjection.

ABSTRACT: The CRISPR/Cas9 system has become a powerful tool for molecular design breeding in livestock such as sheep. However, the efficiency of the Cas9 system combined with zygote microinjection remains suboptimal. In this study, mature sheep oocytes were used for microinjection to assess the impact of various factors on Cas9 editing efficiency. We found that the in vitro maturation efficiency of oocytes is related to environmental factors such as air temperature, pressure, and humidity. Our results indicate that high-efficiency gene editing can be achieved when targeting the SOCS2, DYA, and TBXT, using a microinjection mixture with a concentration of 10 ng/μL Cas9 and sgRNA. By optimizing the injection capillary, we significantly reduced the oocyte invalidation rate post-micr

SUBMITTER: Wang H 

PROVIDER: S-EPMC11817664 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature