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Identification of Endothelial Specific Region in the Intracellular Adhesion Molecule-2 (ICAM2) Promoter of Miniature Pig

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한국동물번식학회 (The Korean Society of Animal Reproduction)
초록

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

목차
ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Cell Culture    Construction of Luciferase Reporter Gene Vectors    Sequence Analysis    Transfections and Analysis of Promoter Activity    Expression Vector Construction of 1 kb Size Promoter    Transfections and Western Blot Analysis    Immnunocytochemistry   RESULT   Cloning of Mini-pig ICAM2 Promoter   DISCUSSION   REFERENCES
저자
  • Hoon Jang(Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea, Functional Genomics, School of Engineering, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea)
  • Won-Gu Jang(Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea)
  • Dong Un Kim(lRe용enerative Medicine Research Center, Korea Research Institute 01 Bioscience and Biotechnology, Da에eon 305-806, Republic 01 Korea)
  • Eun-Jung Kim(lRe용enerative Medicine Research Center, Korea Research Institute 01 Bioscience and Biotechnology, Da에eon 305-806, Republic 01 Korea)
  • Sung Soo Hwang(Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon 441-706, Republic of Korea)
  • Keon Bong Oh(3 Animal Biotechnology Division, National Institute 01 Animal Science, Rural Development Administration, Suwon 441-706, Republic 01 Korea)
  • Jeong-Woong Lee(Regenerative Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea, Functional Genomics, School of Engineering, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea) Corresponding Author