title: Assessment of mRNA splice variants by qRT-PCR.
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.
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, USA.
authors.forename: Ileabett M Echevarria
authors.initials: IM
authors.lastname: Vargas
authors.forename: Pablo E
authors.initials: PE
authors.lastname: Vivas-Mejía
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D052160:Benzothiazoles; D003959:Diamines; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification*; D011804:Quinolines; D012326:RNA Splicing / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D020033:Protein Isoforms; D011804:Quinolines; C098022:SYBR Green I
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