Expression of GFP Gene in the Porcine Preimplantation Embryos after ICSI with DNA/Sperm Complex

서지정보

· 발행기관 : 한국동물번식학회
· 수록지 정보 : Reproductive & developmental biology / 30권 / 2호
· 저자명 : Joo-Hee Han, Sungwoo Kim, Poongyeon Lee, Chun-Gyu Park, Hyun-Gi Lee, Boh-Suk Yang, Ki-Hyeong Rhee, Chang-Hyung Lee, Hoon-Taek Lee, Won-Kyong Chang, Jin-Ki Park

목차

INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

영어초록

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.

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