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Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype
OBJECTIVE: Nonsense mutations account for 5–70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to gene...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Blackwell Publishing Ltd
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4284126/ https://www.ncbi.nlm.nih.gov/pubmed/25574475 http://dx.doi.org/10.1002/acn3.144 |
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author | Bouchelion, Ashleigh Zhang, Zhongjian Li, Yichao Qian, Haohua Mukherjee, Anil B |
author_facet | Bouchelion, Ashleigh Zhang, Zhongjian Li, Yichao Qian, Haohua Mukherjee, Anil B |
author_sort | Bouchelion, Ashleigh |
collection | PubMed |
description | OBJECTIVE: Nonsense mutations account for 5–70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to generate a reliable mouse model of INCL carrying the most common Ppt1 nonsense mutation (c.451C>T) found in the United States patient population to provide a platform for evaluating nonsense suppressors in vivo. METHODS: We knocked-in c.451C>T nonsense mutation in the Ppt1 gene in C57 embryonic stem (ES) cells using a targeting vector in which LoxP flanked the Neo cassette, which was removed from targeted ES cells by electroporating Cre. Two independently targeted ES clones were injected into blastocysts to generate syngenic C57 knock-in mice, obviating the necessity for extensive backcrossing. RESULTS: Generation of Ppt1-KI mice was confirmed by DNA sequencing, which showed the presence of c.451C>T mutation in the Ppt1 gene. These mice are viable and fertile, although they developed spasticity (a “clasping” phenotype) at a median age of 6 months. Autofluorescent storage materials accumulated throughout the brain regions and in visceral organs. Electron microscopic analysis of the brain and the spleen showed granular osmiophilic deposits. Increased neuronal apoptosis was particularly evident in cerebral cortex and abnormal histopathological and electroretinographic (ERG) analyses attested striking retinal degeneration. Progressive deterioration of motor coordination and behavioral parameters continued until eventual death. INTERPRETATION: Our findings show that Ppt1-KI mice reliably recapitulate INCL phenotype providing a platform for testing the efficacy of existing and novel nonsense suppressors in vivo. |
format | Online Article Text |
id | pubmed-4284126 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Blackwell Publishing Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-42841262015-01-08 Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype Bouchelion, Ashleigh Zhang, Zhongjian Li, Yichao Qian, Haohua Mukherjee, Anil B Ann Clin Transl Neurol Research Articles OBJECTIVE: Nonsense mutations account for 5–70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to generate a reliable mouse model of INCL carrying the most common Ppt1 nonsense mutation (c.451C>T) found in the United States patient population to provide a platform for evaluating nonsense suppressors in vivo. METHODS: We knocked-in c.451C>T nonsense mutation in the Ppt1 gene in C57 embryonic stem (ES) cells using a targeting vector in which LoxP flanked the Neo cassette, which was removed from targeted ES cells by electroporating Cre. Two independently targeted ES clones were injected into blastocysts to generate syngenic C57 knock-in mice, obviating the necessity for extensive backcrossing. RESULTS: Generation of Ppt1-KI mice was confirmed by DNA sequencing, which showed the presence of c.451C>T mutation in the Ppt1 gene. These mice are viable and fertile, although they developed spasticity (a “clasping” phenotype) at a median age of 6 months. Autofluorescent storage materials accumulated throughout the brain regions and in visceral organs. Electron microscopic analysis of the brain and the spleen showed granular osmiophilic deposits. Increased neuronal apoptosis was particularly evident in cerebral cortex and abnormal histopathological and electroretinographic (ERG) analyses attested striking retinal degeneration. Progressive deterioration of motor coordination and behavioral parameters continued until eventual death. INTERPRETATION: Our findings show that Ppt1-KI mice reliably recapitulate INCL phenotype providing a platform for testing the efficacy of existing and novel nonsense suppressors in vivo. Blackwell Publishing Ltd 2014-12 2014-11-18 /pmc/articles/PMC4284126/ /pubmed/25574475 http://dx.doi.org/10.1002/acn3.144 Text en © 2014 The Authors. Annals of Clinical and Translational Neurology published by Wiley Periodicals, Inc on behalf of American Neurological Association. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Bouchelion, Ashleigh Zhang, Zhongjian Li, Yichao Qian, Haohua Mukherjee, Anil B Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title | Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title_full | Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title_fullStr | Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title_full_unstemmed | Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title_short | Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype |
title_sort | mice homozygous for c.451c>t mutation in cln1 gene recapitulate incl phenotype |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4284126/ https://www.ncbi.nlm.nih.gov/pubmed/25574475 http://dx.doi.org/10.1002/acn3.144 |
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