Research Resource: Genetic Labeling of Human Islet Alpha Cells.

ABSTRACT: The 2 most abundant human pancreatic islet cell types are insulin-producing ?-cells and glucagon-producing ?-cells. Defined cis-regulatory elements from rodent Insulin genes have permitted genetic labeling of human islet ?-cells, enabling lineage tracing and generation of human ?-cell lines, but analogous elements for genetically labeling human ?-cells with high specificity do not yet exist. To identify genetic elements that specifically direct reporter expression to human ?-cells, we investigated noncoding sequences adjacent to the human GLUCAGON and ARX genes, which are expressed in islet ?-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 ?- and ?-cells, we found that rat glucagon promoter was not specific for human ?-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 ?-cells should enable targeted genetic modification and lineage tracing.

SUBMITTER: Pauerstein PT 

PROVIDER: S-EPMC4792229 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature