1. In vivo Perturb-Seq reveals neuronal and glial abnormalities associated with autism risk genes.
- Author
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Jin X, Simmons SK, Guo A, Shetty AS, Ko M, Nguyen L, Jokhi V, Robinson E, Oyler P, Curry N, Deangeli G, Lodato S, Levin JZ, Regev A, Zhang F, and Arlotta P
- Subjects
- Animals, Ankyrins genetics, Ankyrins metabolism, CRISPR-Cas Systems, DNA-Binding Proteins genetics, Frameshift Mutation, Gene Expression Profiling, Genetic Loci, Humans, Mice, Neuroglia metabolism, Neurons metabolism, Repressor Proteins genetics, Risk, Transcription Factors genetics, Autistic Disorder genetics, Autistic Disorder pathology, Brain abnormalities, Neuroglia pathology, Neurons pathology
- Abstract
The number of disease risk genes and loci identified through human genetic studies far outstrips the capacity to systematically study their functions. We applied a scalable genetic screening approach, in vivo Perturb-Seq, to functionally evaluate 35 autism spectrum disorder/neurodevelopmental delay (ASD/ND) de novo loss-of-function risk genes. Using CRISPR-Cas9, we introduced frameshift mutations in these risk genes in pools, within the developing mouse brain in utero, followed by single-cell RNA-sequencing of perturbed cells in the postnatal brain. We identified cell type-specific and evolutionarily conserved gene modules from both neuronal and glial cell classes. Recurrent gene modules and cell types are affected across this cohort of perturbations, representing key cellular effects across sets of ASD/ND risk genes. In vivo Perturb-Seq allows us to investigate how diverse mutations affect cell types and states in the developing organism., (Copyright © 2020, American Association for the Advancement of Science.)
- Published
- 2020
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