Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer
Understanding the behavior of transgenes introduced into oocyte or embryos is essential for evaluating the methodologies for transgenic animal production. To date, many studies have reported the production of transgenic pig embryos with, however, low efficiency in environment of blastocyst productio...
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The Korean Society of Animal Reproduction and Biotechnology
2015-09-01
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| Series: | Journal of Animal Reproduction and Biotechnology |
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| Online Access: | http://www.e-jarb.org/journal/view.html?uid=1257&vmd=Full |
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| author | Hak-Jae Chung NaRae Son Joo-Hee Han Chun-Gyu Park Kyung-Woon Kim Mi-Ryung Park In-Sul Hwang Jin-Ki Park Gi-Sun Im |
| author_facet | Hak-Jae Chung NaRae Son Joo-Hee Han Chun-Gyu Park Kyung-Woon Kim Mi-Ryung Park In-Sul Hwang Jin-Ki Park Gi-Sun Im |
| author_sort | Hak-Jae Chung |
| collection | DOAJ |
| description | Understanding the behavior of transgenes introduced into oocyte or embryos is essential for evaluating the methodologies for transgenic animal production. To date, many studies have reported the production of transgenic pig embryos with, however, low efficiency in environment of blastocyst production. The aim of present study was to determine the expression and duration of transgene transferred by intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT). Embryos obtained from the ICSI-MGT procedure were analysed for the expression of GFP and then for the transmission of the transgene. Briefly, fresh spermatozoa were bound to exogenous DNA after treatment by Triton X-100 and Lipofectin. When ICSI-MGT was performed using sperm heads with tails removed, the yield of blastocyst (25.3%), treated with Lipofectin (18.8%) and Triton X-100 (19.2%) were observed. Treatments of Lipofectin or Triton X-100 did not further improve the rates of blastocysts. Moreover, the apoptosis rates of embryos were obtained from the control and LIpofectin groups (8.7%, 9.7%, respectively), but were significantly higher in the Triton X-100 group (13.0%). Our results demonstrated that ICSI-MGT caused minimal damage to oocytes that could develop to full term. Moreover, the embryos derived by ICSI-MGT have shown prolonged exogenous DNA expression during preimplantation stage in vivo. However, more efforts will be required to improve the procedures of both sperm treatments cause of high frequency of mosaicisms. |
| first_indexed | 2024-12-20T05:46:31Z |
| format | Article |
| id | doaj.art-4aadf832e1ad499dbfb81b2ef70f8876 |
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| issn | 2671-4639 2671-4663 |
| language | English |
| last_indexed | 2024-12-20T05:46:31Z |
| publishDate | 2015-09-01 |
| publisher | The Korean Society of Animal Reproduction and Biotechnology |
| record_format | Article |
| series | Journal of Animal Reproduction and Biotechnology |
| spelling | doaj.art-4aadf832e1ad499dbfb81b2ef70f88762022-12-21T19:51:18ZengThe Korean Society of Animal Reproduction and BiotechnologyJournal of Animal Reproduction and Biotechnology2671-46392671-46632015-09-0130322522810.12750/JET.2015.30.3.225Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene TransferHak-Jae Chung0NaRae Son1Joo-Hee Han2Chun-Gyu Park3Kyung-Woon Kim4Mi-Ryung Park5 In-Sul Hwang6Jin-Ki Park7Gi-Sun Im8Animal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, KoreaCHO-A Biotechnology Research Institute, CHO-A Pharmaceutical Co., Ltd., Yeo-Ju 469-841, Korea CHO-A Biotechnology Research Institute, CHO-A Pharmaceutical Co., Ltd., Yeo-Ju 469-841, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, Korea Animal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, Korea Dept. of Swine & Poultry Science, Korea National College of Agriculture and Fisheries, Jeonju 560-500, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, Korea Understanding the behavior of transgenes introduced into oocyte or embryos is essential for evaluating the methodologies for transgenic animal production. To date, many studies have reported the production of transgenic pig embryos with, however, low efficiency in environment of blastocyst production. The aim of present study was to determine the expression and duration of transgene transferred by intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT). Embryos obtained from the ICSI-MGT procedure were analysed for the expression of GFP and then for the transmission of the transgene. Briefly, fresh spermatozoa were bound to exogenous DNA after treatment by Triton X-100 and Lipofectin. When ICSI-MGT was performed using sperm heads with tails removed, the yield of blastocyst (25.3%), treated with Lipofectin (18.8%) and Triton X-100 (19.2%) were observed. Treatments of Lipofectin or Triton X-100 did not further improve the rates of blastocysts. Moreover, the apoptosis rates of embryos were obtained from the control and LIpofectin groups (8.7%, 9.7%, respectively), but were significantly higher in the Triton X-100 group (13.0%). Our results demonstrated that ICSI-MGT caused minimal damage to oocytes that could develop to full term. Moreover, the embryos derived by ICSI-MGT have shown prolonged exogenous DNA expression during preimplantation stage in vivo. However, more efforts will be required to improve the procedures of both sperm treatments cause of high frequency of mosaicisms.http://www.e-jarb.org/journal/view.html?uid=1257&vmd=Fulltransgenic pigicsi-mgtgfp expression |
| spellingShingle | Hak-Jae Chung NaRae Son Joo-Hee Han Chun-Gyu Park Kyung-Woon Kim Mi-Ryung Park In-Sul Hwang Jin-Ki Park Gi-Sun Im Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer Journal of Animal Reproduction and Biotechnology transgenic pig icsi-mgt gfp expression |
| title | Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer |
| title_full | Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer |
| title_fullStr | Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer |
| title_full_unstemmed | Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer |
| title_short | Prolonged Expression of Exogenous GFP Gene in the Porcine Embryos generated by Intracytoplasmic Sperm Injection-Mediated Gene Transfer |
| title_sort | prolonged expression of exogenous gfp gene in the porcine embryos generated by intracytoplasmic sperm injection mediated gene transfer |
| topic | transgenic pig icsi-mgt gfp expression |
| url | http://www.e-jarb.org/journal/view.html?uid=1257&vmd=Full |
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