Abstract：
:The availability of regulatory sequences directing tissue-specific expression of transgenes in genetically modified mice and large animals is a prerequisite for the development of adequate models for human diseases. The rat insulin 2 gene (Ins2) promoter, widely used to achieve transgene expression in pancreatic beta-cells of mice, also directs expression to extrapancreatic tissues and performs poorly in isolated pancreatic islets of human, mouse, and pig. To evaluate whether the full 5' untranslated region (UTR) of the porcine insulin gene (INS) confers robust and specific expression in beta-cells we generated an expression cassette containing 1500bp of the porcine INS 5' UTR and the 3' UTR of the bovine growth hormone gene (GH). The cassette was designed to allow easy exchange of the sequences to be expressed and easy removal of the vector backbone from the expression cassette. To evaluate the properties of the cassette, we initially inserted a cDNA encoding human betacellulin, a growth factor known to affect structural and functional parameters of beta-cells. After confirming the functionality and specificity of the construct in vitro, transgenic mouse lines were generated by pronuclear DNA microinjection. Using RT-PCR, immunohistochemistry and immunofluorescence, we show that transgenic mice expressed human betacellulin exclusively in beta-cells. Confirming the proposed insulinotropic effect of betacellulin, transgenic mice showed improved glucose tolerance. We conclude that the newly designed expression cassette containing 1500bp of the porcine insulin promoter 5' UTR confers robust and specific transgene expression to beta-cells in vitro and in transgenic mice.
journal_name
Mol Cell Endocrinoljournal_title
Molecular and cellular endocrinologyauthors
Grzech M,Dahlhoff M,Herbach N,Habermann FA,Renner-Müller I,Wanke R,Flaswinkel H,Wolf E,Schneider MRdoi
10.1016/j.mce.2009.08.001subject
Has Abstractpub_date
2010-02-05 00:00:00pages
219-24issue
1-2eissn
0303-7207issn
1872-8057pii
S0303-7207(09)00407-9journal_volume
315pub_type
杂志文章abstract：:Melatonin has repeatedly been shown to inhibit the proliferation of MCF-7 human breast cancer cells. Previous reports suggest that the actions of melatonin can be mediated either through G-protein coupled membrane receptors or via retinoid orphan receptors (RORalphas). In this study, we demonstrated the expression of ...
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journal_title：Molecular and cellular endocrinology
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journal_title：Molecular and cellular endocrinology
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journal_title：Molecular and cellular endocrinology
pub_type： 杂志文章
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更新日期：2011-01-01 00:00:00
