In cystic fibrosis (CF), transcript analysis and quantification are important for diagnosis, prognosis and also as surrogate markers for some therapies including gene therapy. Classical RNA-based methods require significant expression levels in target samples for appropriate analysis, thus PCR-based methods are evolving towards reliable quantification. Various protocols for the quantitative analysis of CFTR transcripts (including those resulting from splicing variants) are described and discussed here.
Bibliographical noteFunding Information:
The preparation of this review was supported by the EU-European Thematic Network on Cystic Fibrosis and Related Diseases (EU-QLK3-CT-1999-00241) and grants from the Italian Cystic Fibrosis Research Foundation, the Cystic Fibrosis Trust (UK), Vaincre La Mucoviscidose (France), the Portuguese Foundation for Science and Technology (POCTI/1999/MGI/35737; POCTI/MGI/47382/2002) and the EU (“CF-Chip” EU-QLK3-CT-2001-01982).
- Alternative splicing
- Quantitative PCR
- Real-time PCR