earticle

영어

The production of transgenic livestock has the opportunity to significantly improve human health, enhance nutrition, protect the environment, increase animal welfare, and decrease livestock disease. In this study, bovine PN embryos were injected in cytoplasm with three groups (Ⅰ: sgRNA+Ca9 protein+KI_hFGF_GFP vector, Ⅱ: sgRNA+Ca9 mRNA+KI_hFGF_GFP vector and Ⅲ: bCNS_EX3_sgRNA+pBSK_sgRNA+Ca9 protein+ KI_hFGF_GFP vector) at 18~20 h after IVF, and treated 7.5 uM RS-1 (RAD51-stimulatory compound 1) for 24 h after microinjection. The survival rate post day 2 was higher in group Ⅰ (84.4%) than other groups. At day 4, embryo development potential (78.1%) and GFP expression was significantly highest in group Ⅲ (54.5%) among treatment groups (p<0.05). At day 8, while the blastocyst formation was significantly high in group Ⅰ(32.1%) among three groups, the GFP expression embryo rate of developed blastocyst was higher in group Ⅱ (34.8%) and Ⅲ (38.5%) than in group Ⅰ (17.1%). The total injection efficiency of GFP embryo production from gene injected embryos was a little higher in group I (5.5%, 6/109) than in group II (4.1%, 8/194) and group III (3.6%, 5/139). However, the GFP expressed healthy embryos were the more presented in group II than other groups. Also, when the effect of 7.5 uM RS-1 treatment to the injected embryos for 24 h were examined, the survival rate post day 2 was higher in RS-1 treatment group (72.5%) than in control group (67.3%). The GFP expression rate was also higher in RS-1 treatment group (57.6% and 58.3%) than in control group (38.8% and 19.2%) at day 4 and 8 development in vitro. This study demonstrates that it can be helpful to the production of hFGF knock-in bovine embryos using combination of bCNS_EX3_sgRNA+pBSK_sgRNA+Ca9 protein+KI_hFGF_ GFP vector and treatment with RS-1.