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

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 36권 / 3호
ㆍ저자명 : Hoon Jang, Won-Gu Jang, Dong Un Kim, Eun-Jung Kim, Sung Soo Hwang, Keon Bong Oh, Jeong-Woong Lee

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

영어 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.

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