Absence of an Intron Splicing Silencer in Porcine Smn1 Intron 7 Confers Immunity to the Exon Skipping Mutation in Human SMN2

Thomas Doktor, Schrøder Lisbeth, Henriette Skovgaard Andersen, Sabrina Brøner, A Kitewska, C.B. Sørensen, Brage Storstein Andresen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Abstract

Spinal Muscular Atrophy is caused by homozygous loss of SMN1. All patients retain at least one copy of SMN2 which produces an identical protein but at lower levels due to a silent mutation in exon 7 which results in predominant exclusion of the exon. Therapies targeting the splicing of SMN2 exon 7 have been in development for several years, and their efficacy has been measured using either in vitro cellular assays or in vivo small animal models such as mice. In this study we evaluated the potential for constructing a mini-pig animal model by introducing minimal changes in the endogenous porcine Smn1 gene to maintain the native genomic structure and regulation. We found that while a Smn2-like mutation can be introduced in the porcine Smn1 gene and can diminish the function of the ESE, it would not recapitulate the splicing pattern seen in human SMN2 due to absence of a functional ISS immediately downstream of exon 7. We investigated the ISS region and show here that the porcine ISS is inactive due to disruption of a proximal hnRNP A1 binding site, while a distal hnRNP A1 binding site remains functional but is unable to maintain the functionality of the ISS as a whole.

OriginalsprogEngelsk
Artikelnummere98841
TidsskriftPLOS ONE
Vol/bind9
Udgave nummer6
Sider (fra-til)e98841
Antal sider10
ISSN1932-6203
DOI
StatusUdgivet - 3. jun. 2014

Fingeraftryk

Dyk ned i forskningsemnerne om 'Absence of an Intron Splicing Silencer in Porcine Smn1 Intron 7 Confers Immunity to the Exon Skipping Mutation in Human SMN2'. Sammen danner de et unikt fingeraftryk.

Citationsformater