Research Resource: Genetic Labeling of Human Islet Alpha Cells.
Mol Endocrinol
; 30(2): 248-53, 2016 Feb.
Article
em En
| MEDLINE
| ID: mdl-26745668
The 2 most abundant human pancreatic islet cell types are insulin-producing ß-cells and glucagon-producing α-cells. Defined cis-regulatory elements from rodent Insulin genes have permitted genetic labeling of human islet ß-cells, enabling lineage tracing and generation of human ß-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 ß-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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Coloração e Rotulagem
/
Técnicas Genéticas
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Células Secretoras de Insulina
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article