Establishment of Neurodevelopmental Model for Functional Validation of DNM1L Missense Mutations

Abstract

Drp1, encoded by the DNM1L gene, functions as a GTPase in mitochondrial and peroxisomal fission. The advent of whole-exome sequencing has revealed pathogenic mutations affecting DNM1L associated with neurological phenotypes, ranging from mild dominant optic atrophy to neonatal lethality. However, a platform for studying the functionality of DNM1L variants related to neurological symptoms is absent. Here we designed a system to observe the neurodevelopmental phenotype of DNM1L variants. Due to the lethal effect of DNM1L knockout, we introduced the Tet-Off system to maintain wild-type DNM1L levels, while knocking out the endogenous DNM1L gene by CRISPR/Cas9 system in human embryonic stem cells. The effects of each variant are expected to be analyzed regarding Drp1 functions after differentiating into neural progenitor cells and mature neurons. Our work may help to expand the comprehension of the DNM1L-related neurological symptom spectrum, and thus provide clinical insights.