Quantitative methods for the analysis of CFTR transcripts/splicing variants.
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.
Amaral
Margarida D
MD
Department of Chemistry and Biochemistry, University of Lisboa, Lisboa, Portugal. mdamaral@fc.ul.pt
Clarke
Luka A
LA
Ramalho
Anabela S
AS
Beck
Sebastian
S
Broackes-Carter
Fiona
F
Rowntree
Rebecca
R
Mouchel
Nathalie
N
Williams
Sarah H
SH
Harris
Ann
A
Tzetis
Maria
M
Steiner
Bernhard
B
Sanz
Javier
J
Gallati
Sabina
S
Nissim-Rafinifa
Malka
M
Kerem
Batsheva
B
Hefferon
Timothy
T
Cutting
Garry R
GR
Goina
Elisa
E
Pagani
Franco
F
D003550:Cystic Fibrosis / Q000175:diagnosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005821:Genetic Techniques; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic / Q000235:genetics
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator