title: Research Resource: Genetic Labeling of Human Islet Alpha Cells.
abstract: 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.
authors.affiliation: Department of Developmental Biology (P.T.P., K.M.P., H.S.P., J.W., S.K.K.) and Howard Hughes Medical Institute (S.K.K.), Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Philip T
authors.initials: PT
authors.lastname: Pauerstein
authors.affiliation: Department of Developmental Biology (P.T.P., K.M.P., H.S.P., J.W., S.K.K.) and Howard Hughes Medical Institute (S.K.K.), Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Keon Min
authors.initials: KM
authors.lastname: Park
authors.affiliation: Department of Developmental Biology (P.T.P., K.M.P., H.S.P., J.W., S.K.K.) and Howard Hughes Medical Institute (S.K.K.), Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Heshan S
authors.initials: HS
authors.lastname: Peiris
authors.affiliation: Department of Developmental Biology (P.T.P., K.M.P., H.S.P., J.W., S.K.K.) and Howard Hughes Medical Institute (S.K.K.), Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Developmental Biology (P.T.P., K.M.P., H.S.P., J.W., S.K.K.) and Howard Hughes Medical Institute (S.K.K.), Stanford University School of Medicine, Stanford, California 94305.
authors.forename: Seung K
authors.initials: SK
authors.lastname: Kim
mesh_terms: D000818:Animals; D001483:Base Sequence; D056426:Genetic Loci; D005821:Genetic Techniques; D057809:HEK293 Cells; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation; D016513:Mice, SCID; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D013194:Staining and Labeling