| Autor: |
Heydemann, Ahlke, Warming, Soren, Clendenin, Cynthia, Sigrist, Kirsten, Hjorth, J. Peter, Simon, M. Celeste |
| Zdroj: |
Blood; November 2000, Vol. 96 Issue: 9 p3040-3048, 9p |
| Abstrakt: |
The c-fesproto-oncogene encodes a 92-kd protein tyrosine kinase whose expression is restricted largely to myeloid and endothelial cells in adult mammals. A 13.2-kilobase (kb) humanc-fesgenomic fragment was previously shown to containcis-acting element(s) sufficient for a locus control function in bone marrow macrophages. Locus control regions (LCRs) confer transgene expression in mice that is integration site independent, copy number dependent, and similar to endogenous murine messenger RNA levels. To identify sequences required for this LCR,c-festransgenes were analyzed in mice. Myeloid-cell–specific, deoxyribonuclease-I–hypersensitive sites localized to the 3′ boundary of exon 1 and intron 3 are required to confer high-level transgene expression comparable to endogenous c-fes, independent of integration site. We define a minimal LCR element as DNA sequences (nucleotides +28 to +2523 relative to the transcription start site) located within intron 1 to intron 3 of the human locus. When this 2.5-kb DNA fragment was linked to a c-fescomplementary DNA regulated by its own 446–base-pair promoter, integration-site–independent, copy-number–dependent transcription was observed in myeloid cells in transgenic mice. Furthermore, this 2.5-kb cassette directed expression of a heterologous gene (enhanced green fluorescent protein) exclusively in myeloid cells. The c-fesregulatory unit represents a novel reagent for targeting gene expression to macrophages and neutrophils in transgenic mice. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
