Functional analysis by minigene assay of putative splicing variants found in Bardet-Biedl syndrome patients

Journal of Cellular and Molecular Medicine
María Alvarez-SattaDiana Valverde

Abstract

Bardet-Biedl syndrome (BBS) and Alström syndrome (ALMS) are rare diseases belonging to the group of ciliopathies. Although mutational screening studies of BBS/ALMS cohorts have been extensively reported, little is known about the functional effect of those changes. Thus, splicing variants are estimated to represent 15% of disease-causing mutations, and there is growing evidence that many exonic changes are really splicing variants misclassified. In this study, we aimed to analyse for the first time several variants in BBS2, ARL6/BBS3, BBS4 and ALMS1 genes predicted to produce aberrant splicing by minigene assay. We found discordance between bioinformatics analysis and experimental data when comparing wild-type and mutant constructs. Remarkably, we identified nonsense variants presumably resistant to nonsense-mediated decay, even when a premature termination codon would be introduced in the second amino acid (p.(G2*) mutation in ARL6/BBS3 gene). As a whole, we report one of the first functional studies of BBS/ALMS1 variants using minigene assay, trying to elucidate their role in disease. Functional studies of variants identified in BBS and ALMS patients are essential for their proper classification and subsequent genetic counsel...Continue Reading

References

Oct 1, 1997·Journal of Computational Biology : a Journal of Computational Molecular Cell Biology·M G ReeseD Haussler
Apr 23, 2002·Nature Reviews. Genetics·Luca CartegniAdrian R Krainer
Jul 13, 2002·Science·William G FairbrotherChristopher B Burge
Feb 21, 2004·Human Molecular Genetics·Nicholas Katsanis
May 10, 2005·Bioinformatics·Carles Ferrer-CostaModesto Orozco
Jan 9, 2008·The Journal of Cell Biology·Wallace F Marshall
Oct 28, 2008·Human Mutation·Amanda B SpurdleUNKNOWN IARC Unclassified Genetic Variants Working Group
Apr 3, 2009·Nucleic Acids Research·François-Olivier DesmetChristophe Béroud
Aug 4, 2009·EMBO Reports·Diana BaralleEmanuele Buratti
Apr 1, 2010·Nature Methods·Ivan A AdzhubeiShamil R Sunyaev
May 26, 2010·Proceedings of the National Academy of Sciences of the United States of America·Norann A ZaghloulNicholas Katsanis
Dec 16, 2010·European Journal of Human Genetics : EJHG·Ines PereiroPhilip L Beales
Apr 22, 2011·The New England Journal of Medicine·Friedhelm HildebrandtNicholas Katsanis
Jul 14, 2011·Genome Research·Timothy Sterne-WeilerJeremy R Sanford
Sep 1, 2011·Nature Reviews. Genetics·Zuben E Sauna, Chava Kimchi-Sarfaty
Nov 2, 2011·Current Genomics·Jan D MarshallJürgen K Naggert
Dec 6, 2011·Proceedings of the National Academy of Sciences of the United States of America·Qihong ZhangVal C Sheffield
Jun 21, 2012·European Journal of Human Genetics : EJHG·Elizabeth Forsythe, Philip L Beales
Nov 19, 2013·Neuroscience and Biobehavioral Reviews·Lam Son NguyenJozef Gecz
Feb 25, 2014·Cilia·Mohamed H Al-HamedJohn A Sayer
Mar 29, 2014·European Journal of Human Genetics : EJHG·Ane Y SteffensenThomas vO Hansen
Aug 30, 2014·Human Molecular Genetics·Kinga M BujakowskaEric A Pierce
Mar 18, 2015·Molecular and Cellular Biology·Daniel MurphyPeter Stoilov
Apr 8, 2015·Human Mutation·Jan D MarshallJürgen K Naggert
Jun 18, 2015·Journal of Medical Genetics·Sheila Castro-SánchezDiana Valverde
Nov 11, 2015·Human Mutation·Susana IgrejaMargarida D Amaral
Jan 15, 2016·Clinical Genetics·S A KhanS Khan
Oct 30, 2016·Human Molecular Genetics·Elise HeonVal C Sheffield

❮ Previous
Next ❯

Methods Mentioned

BETA
genotyping
PCR
electrophoresis
transfection
antisense oligonucleotides

Software Mentioned

HSF ( Human Splicing Finder
NetGene2 SPLICE ( NNSplice )
ASSEDA ( Automated Splice Site Exon Definition Server )
Rescue ESE ( Enhancer
Pmut
Rescue ESE
SIFT
NNSplice SPLICE
Splice Site Exon Definition Server ‐ ASSEDA
ASSEDA

Related Concepts

Related Feeds

Alstrom Syndrome

Alstrom syndrome is a rare disorder characterized by early obesity, loss of central vision, diabetes mellitus, hearing loss, and short stature. Here is the latest research.