Assessment of mRNA splice variants by qRT-PCR.

Imagen de Pablo Vivas-Mejia
PDF versionPDF version
TítuloAssessment of mRNA splice variants by qRT-PCR.
Publication TypeJournal Article
Year of Publication2013
AutoresVargas, IMEchevarr, Vivas-Mejia, P
JournalMethods Mol Biol
Volume1049
Pagination171-86
Date Published2013
ISSN1940-6029
Palabras claveApoptosis, Female, Gene Expression Regulation, Neoplastic, Humans, Organic Chemicals, Ovarian Neoplasms, Protein Isoforms, Real-Time Polymerase Chain Reaction, RNA Splicing
Abstract

Alternative splicing is an essential process for the generation of protein diversity. The physiological role, cellular localization, and abundance of splice variant products compared to the wild-type protein may be completely different. This is illustrated by the five splice variants of the antiapoptotic protein survivin that are more abundant in cancerous cells compared with normal tissues. Interestingly, some survivin splice variants have been associated with drug resistance. Herein, we describe a SYBR green I-based real-time PCR method to assess the messenger RNA levels of the human survivin splice variants in taxane-sensitive versus taxane-resistant ovarian cancer cells and in human ovarian cancer samples. Furthermore, in this chapter, we describe the quantification of survivin splice variants by real-time quantitative PCR (qPCR) after in vitro and in vivo small interference RNA (siRNA)-mediated silencing of survivin splice variants.

DOI10.1007/978-1-62703-547-7_13
Alternate JournalMethods Mol. Biol.
PubMed ID23913216
PubMed Central IDPMC4060277
Grant ListK22 CA166226 / CA / NCI NIH HHS / United States
R25-GM061838 / GM / NIGMS NIH HHS / United States