研究资源:人胰岛α细胞的遗传标记。

Q Biochemistry, Genetics and Molecular Biology Molecular endocrinology Pub Date : 2016-01-08 DOI:10.1210/me.2015-1220
P. Pauerstein, K. Park, H. Peiris, Jing Wang, Seung K. Kim
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引用次数: 6

摘要

2种最丰富的人类胰岛细胞类型是产生胰岛素的β-细胞和产生胰高血糖素的α-细胞。从啮齿动物胰岛素基因中确定的顺式调控元件允许人类胰岛β细胞的遗传标记,使谱系追踪和人类β细胞系的产生成为可能,但具有高特异性的人类α细胞遗传标记的类似元件尚不存在。为了鉴定特异性指导人α-细胞报告基因表达的遗传元件,我们研究了在胰岛α-细胞中表达的人胰高血糖素和ARX基因附近的非编码序列。将具有高度进化保守性的元件克隆到慢病毒载体中,在人胰岛中直接荧光报告基因表达。根据α-和β-细胞报告基因表达的特异性,我们发现大鼠胰高血糖素启动子对人α-细胞不具有特异性,但据我们所知,人胰高血糖素非翻译区序列的加入大大提高了培养和移植胰岛的标记特异性,其程度在一定程度上未被报道过。这些顺式调控序列在人类α-细胞中的特异性转基因表达应该能够实现靶向遗传修饰和谱系追踪。
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Research Resource: Genetic Labeling of Human Islet Alpha Cells.
The 2 most abundant human pancreatic islet cell types are insulin-producing β-cells and glucagon-producing α-cells. Defined cis-regulatory elements from rodent Insulin genes have permitted genetic labeling of human islet β-cells, enabling lineage tracing and generation of human β-cell lines, but analogous elements for genetically labeling human α-cells with high specificity do not yet exist. To identify genetic elements that specifically direct reporter expression to human α-cells, we investigated noncoding sequences adjacent to the human GLUCAGON and ARX genes, which are expressed in islet α-cells. Elements with high evolutionary conservation were cloned into lentiviral vectors to direct fluorescent reporter expression in primary human islets. Based on the specificity of reporter expression for α- and β-cells, we found that rat glucagon promoter was not specific for human α-cells but that addition of human GLUCAGON untranslated region sequences substantially enhanced specificity of labeling in both cultured and transplanted islets to a degree not previously reported, to our knowledge. Specific transgene expression from these cis-regulatory sequences in human α-cells should enable targeted genetic modification and lineage tracing.
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来源期刊
Molecular endocrinology
Molecular endocrinology 医学-内分泌学与代谢
CiteScore
3.49
自引率
0.00%
发文量
0
审稿时长
12 months
期刊介绍: Molecular Endocrinology provides a forum for papers devoted to describing molecular mechanisms by which hormones and related compounds regulate function. It has quickly achieved a reputation as a high visibility journal with very rapid communication of cutting edge science: the average turnaround time is 28 days from manuscript receipt to first decision, and accepted manuscripts are published online within a week through Rapid Electronic Publication. In the 2008 Journal Citation Report, Molecular Endocrinology is ranked 16th out of 93 journals in the Endocrinology and Metabolism category, with an Impact Factor of 5.389.
期刊最新文献
Editorial Reflections on the Demise of Molecular Endocrinology and the Future of Molecular Hormone Action Research. Origins of the Field of Molecular Endocrinology: A Personal Perspective. Editorial: Reflections on the Impact of Molecular Endocrinology on a Scientific Career. Reflections on the Merger of Molecular Endocrinology and Endocrinology. Editorial: Final Musings on the Impact of Molecular Endocrinology.
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