2 research outputs found
Functional analysis of new human Bardet-Biedl syndrome loci specific variants in the zebrafish model
The multiple genetic approaches available for molecular diagnosis of human diseases have made possible to identify an increasing number of pathogenic genetic changes, particularly with the advent of next generation sequencing (NGS) technologies. However, the main challenge lies in the interpretation of their functional impact, which has resulted in the widespread use of animal models. We describe here the functional modelling of seven BBS loci variants, most of them novel, in zebrafish embryos to validate their in silico prediction of pathogenicity. We show that target knockdown (KD) of known BBS (BBS1, BB5 or BBS6) loci leads to developmental defects commonly associated with ciliopathies, as previously described. These KD pleiotropic phenotypes were rescued by co-injecting human wild type (WT) loci sequence but not with the equivalent mutated mRNAs, providing evidence of the pathogenic effect of these BBS changes. Furthermore, direct assessment of cilia located in Kupfferâs vesicle (KV) showed a reduction of ciliary length associated with all the studied variants, thus confirming a deleterious effect. Taken together, our results seem to prove the pathogenicity of the already classified and unclassified new BBS variants, as well as highlight the usefulness of zebrafish as an animal model for in vivo assays in human ciliopathiesMinisterio de EconomĂa y Competitividad | Ref. AGL2017-89648
Functional analysis of new human Bardet-Biedl syndrome loci specific variants in the zebrafish model
12 pages, 7 figures, 4 tables.-- This article is licensed under a Creative Commons Attribution 4.0 International
LicenseThe multiple genetic approaches available for molecular diagnosis of human diseases have made
possible to identify an increasing number of pathogenic genetic changes, particularly with the advent of
next generation sequencing (NGS) technologies. However, the main challenge lies in the interpretation
of their functional impact, which has resulted in the widespread use of animal models. We describe
here the functional modelling of seven BBS loci variants, most of them novel, in zebrafish embryos to
validate their in silico prediction of pathogenicity. We show that target knockdown (KD) of known BBS
(BBS1, BB5 or BBS6) loci leads to developmental defects commonly associated with ciliopathies, as
previously described. These KD pleiotropic phenotypes were rescued by co-injecting human wild type
(WT) loci sequence but not with the equivalent mutated mRNAs, providing evidence of the pathogenic
effect of these BBS changes. Furthermore, direct assessment of cilia located in Kupfferâs vesicle
(KV) showed a reduction of ciliary length associated with all the studied variants, thus confirming a
deleterious effect. Taken together, our results seem to prove the pathogenicity of the already classified
and unclassified new BBS variants, as well as highlight the usefulness of zebrafish as an animal model
for in vivo assays in human ciliopathiesThis work was funded by the Spanish Economy and Competitiveness Ministry
projects AGL2017-89648P to JR and PSBPeer reviewe