Conditional Estrogen Receptor Knockout Mouse Model for Studying Mammary Tumorigenesis
Annual rept. 1 Jan-31 Dec 2001
CINCINNATI UNIV OH
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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 ERTK 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.
- Medicine and Medical Research