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 from an androgen dependent to androgen independent phenotype. It is hypothesized that the factors that control this splicing "switch" may be involved in the acquisition of androgen independent growth. To accomplish these goals, we are making use of a genetic screening strategy to identify splicing factors that specifically block the splicing of an exon, IIIc, whose inclusion is seen in androgen independent cancers. We have established reporter constructs in which splicing is required in order to restore an open reading frame (ORF)to a green fluorescent protein (GFP) expression plasmid. An adenoviral derived intron has placed in the GFP ORF and been shown to be efficiently spliced to yield GFP expression that can be identified by flow cytometry. We have inserted FGFR2 sequences containing ;This intron and been able to show cell-type specific splicing of exon IIIc. Currently we are further establishing the screen by optimizing exon IIIc splicing in GFP reporter contructs prior to performing cDNA screening.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2001
- Accession Number
- ADA407310
Entities
People
- Russ P. Carstens
Organizations
- University of Pennsylvania