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通过细菌人工染色体重组工程实现CreErT在腺泡细胞中的强大转基因表达。

Robust acinar cell transgene expression of CreErT via BAC recombineering.

作者信息

Ji Baoan, Song Jian, Tsou Lilian, Bi Yan, Gaiser Sebastian, Mortensen Richard, Logsdon Craig

机构信息

Department of Cancer Biology, University of Texas, M.D. Anderson Cancer Center, Houston, Texas 77030, USA.

出版信息

Genesis. 2008 Aug;46(8):390-5. doi: 10.1002/dvg.20411.

Abstract

Pancreatic acinar cells are critical in gastrointestinal physiology and pancreatitis and may be involved in pancreatic cancer. Previously, a short rat pancreatic elastase promoter has been widely utilized to control acinar cell transgene expression. However, this partial sequence does not confer robust and stable expression. In this study, we tested the hypothesis that a transgene employing bacterial-artificial-chromosome (BAC) technology to express a tamoxifen-regulated Cre recombinase from a full-length mouse elastase gene (BAC-Ela-CreErT) would be more robust and stable. When founders were crossed with Rosa26 reporter mice nearly 100% of acini expressed beta-galactosidase after tamoxifen treatment. The expression was specific for pancreatic acinar cells and these characteristics have remained stable for 2 years. However, because of high levels of expression in differentiated acinar cells, this construct is tamoxifen independent in approximately 50% of adult acinar cells. This model of pancreatic acinar specific Cre expression is a powerful tool for future transgenic and knockout studies.

摘要

胰腺腺泡细胞在胃肠生理学和胰腺炎中至关重要,并且可能与胰腺癌有关。此前,一个短的大鼠胰腺弹性蛋白酶启动子已被广泛用于控制腺泡细胞转基因表达。然而,这个部分序列并不能赋予强大而稳定的表达。在本研究中,我们检验了这样一个假设:采用细菌人工染色体(BAC)技术从全长小鼠弹性蛋白酶基因表达他莫昔芬调节的Cre重组酶的转基因(BAC-Ela-CreErT)会更强大且稳定。当奠基者与Rosa26报告基因小鼠杂交时,在给予他莫昔芬处理后,近100%的腺泡表达β-半乳糖苷酶。该表达对胰腺腺泡细胞具有特异性,并且这些特征已稳定保持了2年。然而,由于在分化的腺泡细胞中表达水平较高,该构建体在大约50%的成年腺泡细胞中对他莫昔芬不依赖。这种胰腺腺泡特异性Cre表达模型是未来转基因和基因敲除研究的有力工具。

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