Promoter and Cofactor Requirements for SERM-ER Activity
Abstract
The mechanisms of estrogen-mediated transcription are not completely understood and as such, the roles of Selective Estrogen Receptor Modulators (SERMs), such as tamoxifen are also poorly understood. Our current work is focused on assessing the relative contributions of the specific promoter sequences within estrogen target genes and how they influence the transcriptional activity by different ligands. Specifically, we have generated breast (MCF-7) and endometrial (ECC1) cancer cell lines with a Lox-Luciferase cassette integrated within the chromatin. These clonal cell lines have been screened by Southern blot and FISH to confirm the presence of a single integration site. Different promoter sequences from estrogen regulated genes have been introduced into these donor cell lines via Cre-mediated recombination. To insert these specific promoter sequences into the same site within the chromatin, we generated an insertion vector containing a multiple cloning sites flanked by Lox sites. After integration of the various promoter sequences we used negative selection to generate cell lines that contained the different promoter sequences within the same chromatin setting. These cell lines are currently being assessed for luciferase activity in the presence of different ligands.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2005
- Accession Number
- ADA435797
Entities
People
- Jason S. Carroll
Organizations
- Dana–Farber Cancer Institute