Methods for identifying and quantifying mRNA expression of androgen receptor splicing variants in prostate cancer

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages165-177
Number of pages13
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1443
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
Grant Support: Studies in the Dehm Lab are supported by NCI Grant R01CA174777 (to S.M.D.) and an American Cancer Society Research Scholar Grant RSG-12-031-01-TBE (to S.M.D.).

Publisher Copyright:
© Springer Science+Business Media New York 2016.

Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.

Keywords

  • AR splice variant
  • Absolute quantification
  • Alternative splicing
  • Androgen receptor
  • Castration-resistant
  • Prostate cancer
  • RT-PCR

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