Identification of Alternative Splicing Factors Involved in Prostate Cancer Progression
Abstract
The goal of this project is to identify and characterize proteins factors that regulate splicing of fibroblast growth factor 2 (FGFR2) in prostate cancer cells. A change in the splicing of this transcript has been shown to accompany the transition of model prostate cancers form an androgen dependent to androgen independent phenotype. It is hypothesized that the factor that control this splicing "switch" may be involved in the acquisition of androgen independent growth. To accomplish these goals, we have established a genetic screening strategy to identify splicing factors that regulate FGFR2 splicing though activation of exon IIIb and repression of splicing of exon IIIc. This switch form exon IIIb to exon Ilic splicing has been shown to occur during the transition to androgen independent cancers. We have established robust splicing reporter constructs in which splicing regulation that yields exon Ilib splicing and IIIc repression can be determined by EGFP or mRFP fluorescence. We have established stable cell lines expressing these reporter and are carrying out genetic screens to identify splicing regulatory proteins. We have set up a retroviral cDNA library screening system in order to carry out his screen. Using dual fluorescence to enhance the efficacy of the system, we expect this strategy to be highly effective as a means to identify functionally relevant proteins.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2003
- Accession Number
- ADA420782
Entities
People
- Russ P. Carstens
Organizations
- University of Pennsylvania