Journal
MOLECULAR ENDOCRINOLOGY
Volume 30, Issue 2, Pages 248-253Publisher
OXFORD UNIV PRESS INC
DOI: 10.1210/me.2015-1220
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Funding
- Howard Hughes Medical Institute (HHMI) Medical Fellows program
- Stanford Medical Scientist Training Program
- United States National Institutes of Health (NIH) [F30DK102301]
- Targeted Resource Generation Project from the NIH Beta Cell Biology Consortium, HHMI, JDRF
- Helmsley Charitable Trust
- HL Snyder Medical Foundation
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The 2 most abundant human pancreatic islet cell types are insulin-producing beta-cells and glucagonproducing alpha-cells. Defined cis-regulatory elements from rodent Insulin genes have permitted genetic labeling of human islet beta-cells, enabling lineage tracing and generation of human beta-cell lines, but analogous elements for genetically labeling human alpha-cells with high specificity do not yet exist. To identify genetic elements that specifically direct reporter expression to human alpha-cells, we investigated noncoding sequences adjacent to the human GLUCAGON and ARX genes, which are expressed in islet alpha-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 alpha- and beta-cells, we found that rat glucagon promoter was not specific for human alpha-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 alpha-cells should enable targeted genetic modification and lineage tracing.
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