CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)

Eaton, S. L. et al. (2019) CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease). Scientific Reports, 9, 9891. (doi: 10.1038/s41598-019-45859-9) (PMID:31289301) (PMCID:PMC6616324)

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Abstract

The neuronal ceroid lipofuscinoses (NCLs) are a group of devastating monogenetic lysosomal disorders that affect children and young adults with no cure or effective treatment currently available. One of the more severe infantile forms of the disease (INCL or CLN1 disease) is due to mutations in the palmitoyl-protein thioesterase 1 (PPT1) gene and severely reduces the child’s lifespan to approximately 9 years of age. In order to better translate the human condition than is possible in mice, we sought to produce a large animal model employing CRISPR/Cas9 gene editing technology. Three PPT1 homozygote sheep were generated by insertion of a disease-causing PPT1 (R151X) human mutation into the orthologous sheep locus. This resulted in a morphological, anatomical and biochemical disease phenotype that closely resembles the human condition. The homozygous sheep were found to have significantly reduced PPT1 enzyme activity and accumulate autofluorescent storage material, as is observed in CLN1 patients. Clinical signs included pronounced behavioral deficits as well as motor deficits and complete loss of vision, with a reduced lifespan of 17 ± 1 months at a humanely defined terminal endpoint. Magnetic resonance imaging (MRI) confirmed a significant decrease in motor cortical volume as well as increased ventricular volume corresponding with observed brain atrophy and a profound reduction in brain mass of 30% at necropsy, similar to alterations observed in human patients. In summary, we have generated the first CRISPR/Cas9 gene edited NCL model. This novel sheep model of CLN1 disease develops biochemical, gross morphological and in vivo brain alterations confirming the efficacy of the targeted modification and potential relevance to the human condition.

Item Type:Articles
Additional Information:This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) through BB/J004316/1 and BB/P013732/1 ISP support to The Roslin Institute and by the Roslin Foundation; BioMarin Pharmaceutical Inc. for sheep maintainance; The RS 370 MacDonald Trust, Simons Initiative for the Developing Brain (SIDB) and Euan MacDonald Centre contributed to the MRI investigations in this pilot study. NMR was funded by a Wellcome Trust Integrated Training Fellowship for Veterinarians (096409/Z/11/Z).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Hamer, Miss Kim and Eaton, Sally
Authors: Eaton, S. L., Proudfoot, C., Lillico, S.G., Skehel, P., Kline, R.A., Hamer, K., Rzechorzek, N.M., Clutton, E., Gregson, R., King, T., O'Neill, C.A., Cooper, J.D., Thompson, G., Whitelaw, C.B., and Wishart, T.M.
College/School:College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
Journal Name:Scientific Reports
Publisher:Nature Research
ISSN:2045-2322
ISSN (Online):2045-2322
Published Online:09 July 2019
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Scientific Reports 9:9891
Publisher Policy:Reproduced under a Creative Commons license

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