Minimal Dataset for Characterization of a C9orf72 Knockout Danio rerio Model for ALS and Cross-Species Validation of Therapeutics in Caenorhabditis elegans

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

<h3>Abstract:</h3>

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

<p>
Various <em>Danio rerio</em> (zebrafish) models of <em>C9orf72</em> depletion have been developed to investigate disease mechanisms and the effects of
<em>C9orf72</em> 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 <em>C9orf72</em> knockout model using CRISPR/Cas9. The <em>C9orf72</em> LOF model exhibits, in a generally
dose-dependent manner, increased larval mortality, persistent growth reduction, and motor deficits. Additionally, homozygous <em>C9orf72</em> LOF larvae
displayed mild overbranching of spinal motoneurons.
</p>

<p>
To identify potential therapeutic compounds, we conducted a screen in an established <em>Caenorhabditis elegans</em> (<em>C. elegans</em>) <em>C9orf72</em>
homologue (<em>alfa-1</em>) 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 <em>C9orf72</em> LOF model.
<strong>Pizotifen malate</strong> was found to significantly improve motor deficits in <em>C9orf72</em> LOF zebrafish larvae.
We present a novel zebrafish <em>C9orf72</em> knockout model that exhibits phenotypic differences from depletion models,
providing a valuable tool for in vivo <em>C9orf72</em> research and ALS therapeutic validation. Furthermore, we identify
<strong>pizotifen malate</strong> as a promising compound for further preclinical evaluation.
</p>

<h3>Dataset Information:</h3>

<p>
This dataset includes raw and processed numerical data from key experimental assays in zebrafish (<em>Danio rerio</em>) and
<em>Caenorhabditis elegans</em>. 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 <em>C. elegans</em>, it includes data on swimming activity, paralysis assays, neurodegeneration analysis, and drug screening experiments.
</p>

<p>
This dataset represents the minimal dataset required for reproducibility in accordance with <em>PLOS Genetics</em> 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.
</p>