Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2.
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 h...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2014-01-01
|
Series: | PLoS ONE |
Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24892836/pdf/?tool=EBI |
id |
doaj-c49b99396dc74de591f148dc0842cbe3 |
---|---|
record_format |
Article |
spelling |
doaj-c49b99396dc74de591f148dc0842cbe32021-03-04T09:21:45ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0196e9884110.1371/journal.pone.0098841Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2.Thomas Koed DoktorLisbeth Dahl SchrøderHenriette Skovgaard AndersenSabrina BrønerAnna KitewskaCharlotte Brandt SørensenBrage Storstein AndresenSpinal 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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24892836/pdf/?tool=EBI |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Thomas Koed Doktor Lisbeth Dahl Schrøder Henriette Skovgaard Andersen Sabrina Brøner Anna Kitewska Charlotte Brandt Sørensen Brage Storstein Andresen |
spellingShingle |
Thomas Koed Doktor Lisbeth Dahl Schrøder Henriette Skovgaard Andersen Sabrina Brøner Anna Kitewska Charlotte Brandt Sørensen Brage Storstein Andresen Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. PLoS ONE |
author_facet |
Thomas Koed Doktor Lisbeth Dahl Schrøder Henriette Skovgaard Andersen Sabrina Brøner Anna Kitewska Charlotte Brandt Sørensen Brage Storstein Andresen |
author_sort |
Thomas Koed Doktor |
title |
Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. |
title_short |
Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. |
title_full |
Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. |
title_fullStr |
Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. |
title_full_unstemmed |
Absence of an intron splicing silencer in porcine Smn1 intron 7 confers immunity to the exon skipping mutation in human SMN2. |
title_sort |
absence of an intron splicing silencer in porcine smn1 intron 7 confers immunity to the exon skipping mutation in human smn2. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2014-01-01 |
description |
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. |
url |
https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24892836/pdf/?tool=EBI |
work_keys_str_mv |
AT thomaskoeddoktor absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT lisbethdahlschrøder absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT henrietteskovgaardandersen absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT sabrinabrøner absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT annakitewska absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT charlottebrandtsørensen absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 AT bragestorsteinandresen absenceofanintronsplicingsilencerinporcinesmn1intron7confersimmunitytotheexonskippingmutationinhumansmn2 |
_version_ |
1714807233111392256 |