PMID- 31289301
OWN - NLM
STAT- MEDLINE
DCOM- 20201027
LR  - 20230107
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Jul 9
TI  - CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid 
      lipofuscinosis (CLN1 disease).
PG  - 9891
LID - 10.1038/s41598-019-45859-9 [doi]
LID - 9891
AB  - 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.
FAU - Eaton, S L
AU  - Eaton SL
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - Proudfoot, C
AU  - Proudfoot C
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - Lillico, S G
AU  - Lillico SG
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - Skehel, P
AU  - Skehel P
AD  - Centre for Discovery Brain Science, University of Edinburgh, Hugh Robson 
      Building, Edinburgh, UK.
AD  - Euan MacDonald Centre for Motor Neurone Disease Research, University of 
      Edinburgh, Edinburgh, UK.
FAU - Kline, R A
AU  - Kline RA
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - Hamer, K
AU  - Hamer K
AUID- ORCID: 0000-0002-6142-5366
AD  - Royal (Dick) School of Veterinary Studies, The University of Edinburgh, 
      Edinburgh, UK.
FAU - Rzechorzek, N M
AU  - Rzechorzek NM
AUID- ORCID: 0000-0003-3209-5019
AD  - Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor's 
      Building, 49 Little France Crescent, Edinburgh, UK.
AD  - Royal (Dick) School of Veterinary Studies, The University of Edinburgh, 
      Edinburgh, UK.
FAU - Clutton, E
AU  - Clutton E
AD  - Wellcome Trust Critical Care Laboratory for Large Animals, Roslin Institute, 
      Easter Bush, Edinburgh, UK.
FAU - Gregson, R
AU  - Gregson R
AD  - Wellcome Trust Critical Care Laboratory for Large Animals, Roslin Institute, 
      Easter Bush, Edinburgh, UK.
FAU - King, T
AU  - King T
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - O'Neill, C A
AU  - O'Neill CA
AD  - BioMarin Pharmaceutical Inc, San Rafael, CA, USA.
FAU - Cooper, J D
AU  - Cooper JD
AD  - Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, David 
      Geffen School of Medicine, UCLA, Torrance, CA, USA.
AD  - Department of Pediatrics, Washington University School of Medicine, St Louis, MO, 
      USA.
FAU - Thompson, G
AU  - Thompson G
AD  - Centre for Discovery Brain Science, University of Edinburgh, Hugh Robson 
      Building, Edinburgh, UK.
FAU - Whitelaw, C B
AU  - Whitelaw CB
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK.
FAU - Wishart, T M
AU  - Wishart TM
AD  - The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of 
      Edinburgh, Edinburgh, UK. T.M.Wishart@ed.ac.uk.
AD  - Euan MacDonald Centre for Motor Neurone Disease Research, University of 
      Edinburgh, Edinburgh, UK. T.M.Wishart@ed.ac.uk.
LA  - eng
GR  - 096409/Z/11/Z/WT_/Wellcome Trust/United Kingdom
GR  - BBS/E/D/20251969/BB_/Biotechnology and Biological Sciences Research 
      Council/United Kingdom
GR  - BB/P013732/1/RCUK | Biotechnology and Biological Sciences Research Council 
      (BBSRC)/International
GR  - MC_EX_MR/S022023/1/MRC_/Medical Research Council/United Kingdom
GR  - WT_/Wellcome Trust/United Kingdom
GR  - BB/J004316/1/RCUK | Biotechnology and Biological Sciences Research Council 
      (BBSRC)/International
GR  - MR/S022023/1/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190709
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - EC 3.1.2.- (Thiolester Hydrolases)
RN  - EC 3.1.2.22 (palmitoyl-protein thioesterase)
SB  - IM
MH  - Animals
MH  - *CRISPR-Cas Systems
MH  - *Disease Models, Animal
MH  - Female
MH  - Male
MH  - *Mutation
MH  - Neuronal Ceroid-Lipofuscinoses/genetics/metabolism/*pathology
MH  - *Phenotype
MH  - Sheep
MH  - Thiolester Hydrolases/*antagonists & inhibitors/genetics
PMC - PMC6616324
COIS- O'Neill CA is an employee of BioMarin Pharmaceutical Inc., San Rafael, CA, USA. 
      TMW is an academic Editor for Scientific Reports.
EDAT- 2019/07/11 06:00
MHDA- 2020/10/28 06:00
PMCR- 2019/07/09
CRDT- 2019/07/11 06:00
PHST- 2018/12/19 00:00 [received]
PHST- 2019/06/12 00:00 [accepted]
PHST- 2019/07/11 06:00 [entrez]
PHST- 2019/07/11 06:00 [pubmed]
PHST- 2020/10/28 06:00 [medline]
PHST- 2019/07/09 00:00 [pmc-release]
AID - 10.1038/s41598-019-45859-9 [pii]
AID - 45859 [pii]
AID - 10.1038/s41598-019-45859-9 [doi]
PST - epublish
SO  - Sci Rep. 2019 Jul 9;9(1):9891. doi: 10.1038/s41598-019-45859-9.