Conditional Estrogen Receptor Knockout Mouse Model for Studying Mammary Tumorigenesis
Abstract
We have finished the construction of the ER-alpha targeting vector and produced ES cell lines. The details of which are provided in the body of this progress report. The KO construct was electroporated into J-1 ES cells. 120 single clones were obtained after selecting the transfected cells with G418. The ES clone genomic DNA is being screened by PCR with ERKO 3' primers to detect the 5.2 kb targeted ER-alpha allele. The targeted allele will be confirmed with ER/TK 3'-Neo primers, ERKO 5'-Neo primers and Southern blot with the 5' probe. The confirmed positive clone will be treated with Cre recombinase to delete the TK-Neo cassette. Following are the highlights of the progress report. (1) Establishment of breast cancer cell lines expressing different ERs. (2) Identification of the target genes of estrogen and BPA in breast cancer cell line expressing only ER-alpha. (3) Contribution the F-domain of ER alpha in the estrogen-dependent recruitment of coactivators. Construction of the ER-alpha targeting vector and generation of ES cell lines for generating conditional ER-alpha KO mice.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2002
- Accession Number
- ADA402348
Entities
People
- Sohaib A. Khan
Organizations
- University of Cincinnati