Functional variants of CFAP410 affect the DNA damage response leading to Motor neuron degeneration – implications for ALS

Ross Ferguson; Vasanta Subramanian

doi:10.1016/j.isci.2025.113338

Functional variants of CFAP410 affect the DNA damage response leading to Motor neuron degeneration – implications for ALS

Ross Ferguson, Vasanta Subramanian

Department of Life Sciences

University College London

Research output: Contribution to journal › Article › peer-review

Abstract

Mutations in CFAP410, a basal body protein known to be required for the formation of primary cilia, have been identified as risk modifiers in amyotrophic lateral sclerosis (ALS), a devastating late onset neurodegenerative disorder with poor prognosis. CFAP410 is also implicated in the DNA damage response and interacts with Nek1, which has been shown to be mutated in ALS. Herein, we investigated the effect of knocking in an HA epitope tag and functional mutations into the endogenous Cfap410 gene by gene editing in mouse embryonic stem cells (mESCs). We show that primary cilia in these edited mESCs, as well as in the neural progenitors and neurons differentiated from them do not exhibit any significant difference in frequency. However, ESCs, neural progenitors, and neurons with knock-in Cfap410 variants are more susceptible to DNA damage and exhibit impaired interaction with Nek1. Our findings point to DNA damage as a convergent pathway leading to ALS.

Original language	English
Article number	113338
Journal	iScience
Volume	28
Issue number	9
Early online date	9 Aug 2025
DOIs	https://doi.org/10.1016/j.isci.2025.113338
Publication status	Published - 19 Sept 2025

Data Availability Statement

• Data: Original western blot images and PCR gel images have been deposited at Mendeley and are publicly available as of the date of publication. The DOI is listed in the key resources table. Microscopy data reported in this article will be shared by the lead contact upon request.
• Code: This article does not report a high-throughput dataset nor original code
• Additional information: Any additional information required to re-analyze the data reported in this article is available from the lead contact upon request.

Funding

We would like to acknowledge T. Yamomoto for the Multiplex CRISPR-Cas9 Assembly System Kit and S. Park for the pDisplay plasmid obtained from Addgene. We would like to thank Allan Bradley for the plasmid pPB-CAG.OSKML-pudtk and Shankar Srinivas for the pCagTag plasmid. The following antibodies were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242: 2H3 developed by Jessell, T.M. & Dodd, J. (HHMI/Columbia University), 40.2D6 developed by Jessell, T.M. & Brenner-Morton, S. (HHMI/Columbia University) and Rat401 (Nestin) developed by Hockfield, S. (Yale University School of Medicine). The research funded by a project grant from the Motor Neurone Disease Association (Project Grant 866-791 ) to VS.

Funders	Funder number
National Institute of Child Health and Human Development
Yale University
National Institutes of Health
Howard Hughes Medical Institute
Motor Neurone Disease Association	866-791
Columbia University	Rat401

Keywords

Cell biology
Molecular biology
Neuroscience

ASJC Scopus subject areas

General

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10.1016/j.isci.2025.113338Licence: CC BY

Cite this

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abstract = "Mutations in CFAP410, a basal body protein known to be required for the formation of primary cilia, have been identified as risk modifiers in amyotrophic lateral sclerosis (ALS), a devastating late onset neurodegenerative disorder with poor prognosis. CFAP410 is also implicated in the DNA damage response and interacts with Nek1, which has been shown to be mutated in ALS. Herein, we investigated the effect of knocking in an HA epitope tag and functional mutations into the endogenous Cfap410 gene by gene editing in mouse embryonic stem cells (mESCs). We show that primary cilia in these edited mESCs, as well as in the neural progenitors and neurons differentiated from them do not exhibit any significant difference in frequency. However, ESCs, neural progenitors, and neurons with knock-in Cfap410 variants are more susceptible to DNA damage and exhibit impaired interaction with Nek1. Our findings point to DNA damage as a convergent pathway leading to ALS.",

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AB - Mutations in CFAP410, a basal body protein known to be required for the formation of primary cilia, have been identified as risk modifiers in amyotrophic lateral sclerosis (ALS), a devastating late onset neurodegenerative disorder with poor prognosis. CFAP410 is also implicated in the DNA damage response and interacts with Nek1, which has been shown to be mutated in ALS. Herein, we investigated the effect of knocking in an HA epitope tag and functional mutations into the endogenous Cfap410 gene by gene editing in mouse embryonic stem cells (mESCs). We show that primary cilia in these edited mESCs, as well as in the neural progenitors and neurons differentiated from them do not exhibit any significant difference in frequency. However, ESCs, neural progenitors, and neurons with knock-in Cfap410 variants are more susceptible to DNA damage and exhibit impaired interaction with Nek1. Our findings point to DNA damage as a convergent pathway leading to ALS.

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Functional variants of CFAP410 affect the DNA damage response leading to Motor neuron degeneration – implications for ALS

Abstract

Data Availability Statement

Funding

Keywords

ASJC Scopus subject areas

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