Gene expression imaging by enzymatic catalysis of a fluorescent probe via membrane-anchored β-glucuronidase

Y. C. Su, K. H. Chuang, Y. M. Wang, C. M. Cheng, S. R. Lin, J. Y. Wang, J. J. Hwang, B. M. Chen, K. C. Chen, S. Roffler, T. L. Cheng

研究成果: 雜誌貢獻文章同行評審

25 引文 斯高帕斯(Scopus)

摘要

Development of nonimmunogenic and specific reporter genes to monitor gene expression in vivo is important for the optimization of gene therapy protocols. We developed a membrane-anchored form of mouse β-glucuronidase (mβG) as a reporter gene to hydrolyze a nonfluorescent glucuronide probe (fluorescein di-β-D-glucuronide, (FDGlcU) to a highly fluorescent reporter to assess the location and persistence of gene expression. A functional β-glucuronidase (βG) was stably expressed on the surface of murine CT26 colon adenocarcinoma cells where it selectively hydrolyzed the cell-impermeable FDGlcU probe. FDGlcU was also preferentially converted to fluorescent probe by (βG) on CT26 tumors. The fluorescent intensity in βG-expressing CT26 tumors was 240 times greater than the intensity in control tumors. Selective imaging of gene expression was also observed after intratumoral injection of adenoviral βG vector into carcinoma xenografts. Importantly, mβG did not induce an antibody response after hydrodynamic plasmid immunization of Balbβc mice, indicating that the reporter gene product displayed low immunogenicity. A membrane-anchored form of human βG also allowed in vivo imaging, demonstrating that human βG can be employed for imaging. This imaging system therefore, displays good selectivity with low immunogenicity and may help assess the location, magnitude and duration of gene expression in living animals and humans.

原文英語
頁(從 - 到)565-574
頁數10
期刊Gene Therapy
14
發行號7
DOIs
出版狀態已發佈 - 四月 2007
對外發佈

ASJC Scopus subject areas

  • 遺傳學

指紋

深入研究「Gene expression imaging by enzymatic catalysis of a fluorescent probe via membrane-anchored β-glucuronidase」主題。共同形成了獨特的指紋。

引用此