title: Methods for Identifying and Quantifying mRNA Expression of Androgen Receptor Splicing Variants in Prostate Cancer.
abstract: Constitutively active androgen receptor (AR) variants (AR-Vs) lacking the AR ligand-binding domain have been identified as drivers of prostate cancer resistance to AR-targeted therapies. A definitive understanding of the role and origin of AR-Vs in the natural history of prostate cancer progression requires cataloging the entire spectrum of AR-Vs expressed in prostate cancer, as well as accurate determination of their expression levels relative to full-length AR in clinical tissues and models of progression. Exon constituency differences at the 3' terminus of mRNAs encoding AR-Vs compared with mRNAs encoding full-length AR can be exploited for discovery and quantification-based experiments. Here, we provide methodological details for 3' rapid amplification of cDNA ends (3' RACE) and absolute quantitative RT-PCR, which are cost-effective approaches for identifying new AR-Vs and quantifying their absolute expression levels in conjunction with full-length AR in RNA samples derived from various sources.
authors.affiliation: Masonic Cancer Center and Department of Laboratory Medicine and Pathology, University of Minnesota Twin Cities, Mayo Mail Code 806, 420 Delaware Street SE, Minneapolis, MN, USA.
authors.forename: Yingming
authors.initials: Y
authors.lastname: Li
authors.affiliation: Masonic Cancer Center and Department of Laboratory Medicine and Pathology, University of Minnesota Twin Cities, Mayo Mail Code 806, 420 Delaware Street SE, Minneapolis, MN, USA. dehm@umn.edu.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Dehm
mesh_terms: D017398:Alternative Splicing; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D011944:Receptors, Androgen / Q000235:genetics*; D014407:Tumor Cells, Cultured
chemical_list: C410798:AR protein, human; D012333:RNA, Messenger; D011944:Receptors, Androgen
keywords: AR splice variant; Absolute quantification; Alternative splicing; Androgen receptor; Castration-resistant; Prostate cancer; RT-PCR