pAR-IRES-EGFP Plasmid

UNMET NEED
The androgen receptor (AR) is a widely expressed ligand-activated transcription factor which mediates androgen signaling by binding to androgen response elements (AREs) in normal tissue and prostate cancer (PCa). Within tumors, the amount of AR plays a crucial role in determining cell growth, resistance to therapy and progression to fatal castrate recurrent PCa in which prostate cells appear to become independent of androgenic steroids. Despite the pivotal role of the AR in male development and fertility and all stages of PCa development, the mechanisms governing AR expression remain poorly understood.

FEATURES
To study the biological function of the natural AR, Johns Hopkins researchers utilize constructs that code for the full-length AR cDNA. AR negative PC-3 human prostate cancer cells were transfected with a plasmid containing the full length coding sequence of AR without its 5’- or 3’-untranslated regions (UTRs) (i.e., pSG5-AR) which inhibited cell growth.
To address the issue of cell growth, the researchers constructed a new expression vector for the AR, pAR-IRES-EGFP, that contains full-length 5'-UTR which includes the identified translation regulatory regions, the full length coding sequence and the partial 3'-UTR, which includes the identified post-transcriptional regulatory regions. When PC-3 cells were transfected with the pAR-IRES-EGFP vector, it was found that transgenic AR protein expression was not growth inhibitory until ligand was added.

STAGE OF DEVELOPMENT
Tangible material available that can
1) Detect the expression of AR-IRES-EGFP transcript by observing fluorescence in living cells.
2) Clones express a comparable level of AR protein to prostate cancer cells expressing endogenous AR protein.

DISEASE INDICATION
Prostate Cancer

ASSOCIATED PUBLICATIONS
■  The Prostate 61_299-304 (2004). ■




 

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