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.
Department of Chemistry and Biochemistry, University of Lisboa, Lisboa, Portugal. mdamaral@fc.ul.pt
Margarida D
MD
Amaral
Luka A
LA
Clarke
Anabela S
AS
Ramalho
Sebastian
S
Beck
Fiona
F
Broackes-Carter
Rebecca
R
Rowntree
Nathalie
N
Mouchel
Sarah H
SH
Williams
Ann
A
Harris
Maria
M
Tzetis
Bernhard
B
Steiner
Javier
J
Sanz
Sabina
S
Gallati
Malka
M
Nissim-Rafinifa
Batsheva
B
Kerem
Timothy
T
Hefferon
Garry R
GR
Cutting
Elisa
E
Goina
Franco
F
Pagani
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