Efficient, glucose responsive and islet-specific transgene expression by a modified rat insulin promoter.
Efficient, glucose responsive and islet-specific transgene expression by a modified rat insulin promoter.
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本研究旨在提高大鼠胰岛素启动子 (RIP) 的效率和胰岛特异性。制备不同长度的大鼠胰岛素启动子并进行体外测试，以驱动 INS1 细胞、α 细胞、腺泡细胞、导管细胞和成纤维细胞中的荧光素酶报告基因表达。 CMV启动子用作阳性对照。此外，通过超声靶向微泡破坏（UTMD）将 DsRed 报告基因体内施用至大鼠胰腺。 UTMD 后使用共聚焦显微镜检测胰腺内 DsRed 的存在和分布。修饰的 RIP3.1 启动子在 INS-1 细胞中包含转录起始位点后的胰岛素基因部分，其活性比全长 RIP 启动子或 CMV 启动子高 5 倍。 RIP3.1 在体外受葡萄糖水平和各种胰岛转录因子调节，并在 α 细胞中表现出活性，但在外分泌细胞中不表现出活性。 RIP3.1-DsRed的体内递送导致DsRed蛋白在β细胞中表达，并且在正常葡萄糖条件下在较小程度上在α细胞中表达。在 RIP3.1 下，外分泌胰腺中不存在 DsRed 信号。修饰的大鼠胰岛素启动子 RIP3.1 可有效且特异性地将基因表达引导至内分泌胰腺。
This study was done to improve efficiency and islet specificity of the rat insulin promoter (RIP). Various rat insulin promoter lengths were prepared and tested in vitro to drive luciferase reporter gene expression in INS1-cells, alpha-cells, acinar cells, ductal cells, and fibroblasts. The CMV promoter was used as a positive control. In addition, the DsRed reporter gene was administered in vivo to rat pancreas by ultrasound-targeted microbubble destruction (UTMD). Confocal microscopy was used to detect the presence and distribution of DsRed within the pancreas after UTMD. A modified RIP3.1 promoter, which includes portions of the insulin gene after its transcription start site is 5-fold more active in INS-1 cells than the full length RIP promoter or the CMV promoter. RIP3.1 is regulated by glucose level and various islet transcription factors in vitro, and exhibits activity in alpha-cells, but not exocrine cells. In vivo delivery of RIP3.1-DsRed resulted in expression of DsRed protein in beta-cells, and to a lesser extent alpha cells under normal glucose conditions. No DsRed signal was present in exocrine pancreas under RIP3.1. A modified rat insulin promoter, RIP3.1, efficiently and specifically directs gene expression to endocrine pancreas.
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影响因子：
4.1
作者：
BOAM, DSW;DOCHERTY, K
通讯作者：
DOCHERTY, K
影响因子：
5.1
作者：
Chen, S.;Ding, J.;Grayburn, P. A.
通讯作者：
Grayburn, P. A.
影响因子：
56.9
作者：
EDLUND, T;WALKER, MD;RUTTER, WJ
通讯作者：
RUTTER, WJ
影响因子：
5.1
作者：
McCarty, DM;Fu, H;Samulski, RJ
通讯作者：
Samulski, RJ
DOI：
10.1073/pnas.89.23.11498
发表时间：
1992-12-01
影响因子：
11.1
作者：
KENNEDY, GC;RUTTER, WJ
通讯作者：
RUTTER, WJ