Höfundar:
Katrin Möller, Juan Ouyang, Kaan Okay, Kasper Hansen, Hans Tómas Björnsson
Introduction: The epigenetic machinery (EM) is a large group of proteins, that read, write or erase marks on DNA and histone tails, thereby regulating both chromatin structure and gene expression. Many of these genes are highly intolerant to mutation and uniformly cause intellectual disability in heterozygous carriers. It is therefore likely that EM components play a critical role in neuronal development, but exactly how is unknown. This project aims to identify if disease-causing EM genes share a transcriptional signature during neuronal development that could explain the overlapping phenotypes among distinct disorders.
Methods: We will systematically knock out 60 EM genes in primary neuronal progenitors (NPCs) using CRISPR/Cas9, differentiate them into functional neurons and use single-cell RNA sequencing to evaluate the transcriptional changes. Overlapping targets will be further explored and validated.
Results: We have successfully established a neurodevelopmental model by isolating NPCs and differentiating them into functional neurons in vitro. We have designed and verified our knock-out system by testing several of the target genes, including Ep300, Kmt2a and Chd1. By targeting NPCs with a comprehensive pool of the 60 guide RNAs we are currently evaluating how individual EM genes affect proliferation and differentiation in our model.
Conclusions: Our results will establish a valuable transcriptome database of multiple individual EM gene knockouts in a single system, modeling neurodevelopment, and yield overlapping downstream gene targets. Our group is currently working on treatments for Kabuki syndrome, one of the EM-disorders, and we hope our findings can identify common targetable pathways for intellectual disability.