DOI: 10.5683/sp3/rtgsag
title: Minimal Dataset for Characterization of a C9orf72 Knockout Danio rerio Model for ALS and Cross-Species Validation of Therapeutics in Caenorhabditis elegans

description: &lt;p&gt;
 This dataset is associated with the manuscript 
 &lt;strong&gt;"Characterization of a C9orf72 Knockout Danio rerio Model for ALS and Cross-Species Validation of Potential Therapeutics Screened in Caenorhabditis elegans"&lt;/strong&gt;, 
 currently under review at &lt;em&gt;PLOS Genetics&lt;/em&gt;. It represents the minimal dataset required for reproducibility under 
 &lt;em&gt;PLOS Genetics&lt;/em&gt; data-sharing policies.
 &lt;/p&gt;

 &lt;h3&gt;Abstract:&lt;/h3&gt;

 &lt;p&gt;
 Intronic hexanucleotide repeat expansions in the &lt;em&gt;C9orf72&lt;/em&gt; gene are the most common genetic cause of the neurodegenerative diseases 
 amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. This expansion reduces &lt;em&gt;C9orf72&lt;/em&gt; expression in affected patients, 
 implicating loss of &lt;em&gt;C9orf72&lt;/em&gt; function (LOF) as a pathogenic mechanism.
 &lt;/p&gt;

 &lt;p&gt;
 Various &lt;em&gt;Danio rerio&lt;/em&gt; (zebrafish) models of &lt;em&gt;C9orf72&lt;/em&gt; depletion have been developed to investigate disease mechanisms and the effects of 
 &lt;em&gt;C9orf72&lt;/em&gt; LOF. However, there are inconsistencies in reported phenotypes, and most have yet to be validated in stable germline ablation models.
 To address this, we generated a zebrafish &lt;em&gt;C9orf72&lt;/em&gt; knockout model using CRISPR/Cas9. The &lt;em&gt;C9orf72&lt;/em&gt; LOF model exhibits, in a generally 
 dose-dependent manner, increased larval mortality, persistent growth reduction, and motor deficits. Additionally, homozygous &lt;em&gt;C9orf72&lt;/em&gt; LOF larvae 
 displayed mild overbranching of spinal motoneurons.
 &lt;/p&gt;

 &lt;p&gt;
 To identify potential therapeutic compounds, we conducted a screen in an established &lt;em&gt;Caenorhabditis elegans&lt;/em&gt; (&lt;em&gt;C. elegans&lt;/em&gt;) &lt;em&gt;C9orf72&lt;/em&gt; 
 homologue (&lt;em&gt;alfa-1&lt;/em&gt;) LOF model, identifying 12 compounds that improved the motility, neurodegeneration, and paralysis phenotypes. 
 Prompted by the shared motor phenotype, 2 of those compounds were tested in our zebrafish &lt;em&gt;C9orf72&lt;/em&gt; LOF model. 
 &lt;strong&gt;Pizotifen malate&lt;/strong&gt; was found to significantly improve motor deficits in &lt;em&gt;C9orf72&lt;/em&gt; LOF zebrafish larvae. 
 We present a novel zebrafish &lt;em&gt;C9orf72&lt;/em&gt; knockout model that exhibits phenotypic differences from depletion models, 
 providing a valuable tool for in vivo &lt;em&gt;C9orf72&lt;/em&gt; research and ALS therapeutic validation. Furthermore, we identify 
 &lt;strong&gt;pizotifen malate&lt;/strong&gt; as a promising compound for further preclinical evaluation.
 &lt;/p&gt;

 &lt;h3&gt;Dataset Information:&lt;/h3&gt;

 &lt;p&gt;
 This dataset includes raw and processed numerical data from key experimental assays in zebrafish (&lt;em&gt;Danio rerio&lt;/em&gt;) and 
 &lt;em&gt;Caenorhabditis elegans&lt;/em&gt;. In zebrafish, it contains data from qPCR, sequencing, western blots, survival assays, motor activity tracking, 
 spinal motor neuron morphology analysis, neuromuscular junction integrity evaluation, and drug screening experiments. 
 For &lt;em&gt;C. elegans&lt;/em&gt;, it includes data on swimming activity, paralysis assays, neurodegeneration analysis, and drug screening experiments.
 &lt;/p&gt;

 &lt;p&gt;
 This dataset represents the minimal dataset required for reproducibility in accordance with &lt;em&gt;PLOS Genetics&lt;/em&gt; guidelines and provides 
 all necessary numerical data to replicate the study’s findings. Where relevant, sample raw files are included to ensure data provenance and reproducibility.
 &lt;/p&gt;
description: This dataset is associated with a manuscript currently under review at PLOS Genetics. 

The README file in the root directory provides general context for the dataset. 

The Main_Metadata_README file contains experimental details for all sub-datasets, along with most of their relevant metadata.
description: ImageJ, v1.53v and version 1.53t
description: Imaris (Oxford Instruments, Switzerland), v8.1.1
description: Volocity (PerkinElmer, UK), 4.1.0 (Build 179)
description: ZEN Blue edition (Carl Zeiss Microscopy), Germany), 2.6.7
description: SnapGene (Dotmatics, USA), various (v4.2.1.1 to 8.0)