P. Pauerstein, K. Park, H. Peiris, Jing Wang, Seung K. Kim
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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.
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.