Single-cell brain organoid screening identifies developmental defects in autism

Nature. 2023 Sep;621(7978):373-380. doi: 10.1038/s41586-023-06473-y. Epub 2023 Sep 13.


The development of the human brain involves unique processes (not observed in many other species) that can contribute to neurodevelopmental disorders1-4. Cerebral organoids enable the study of neurodevelopmental disorders in a human context. We have developed the CRISPR-human organoids-single-cell RNA sequencing (CHOOSE) system, which uses verified pairs of guide RNAs, inducible CRISPR-Cas9-based genetic disruption and single-cell transcriptomics for pooled loss-of-function screening in mosaic organoids. Here we show that perturbation of 36 high-risk autism spectrum disorder genes related to transcriptional regulation uncovers their effects on cell fate determination. We find that dorsal intermediate progenitors, ventral progenitors and upper-layer excitatory neurons are among the most vulnerable cell types. We construct a developmental gene regulatory network of cerebral organoids from single-cell transcriptomes and chromatin modalities and identify autism spectrum disorder-associated and perturbation-enriched regulatory modules. Perturbing members of the BRG1/BRM-associated factor (BAF) chromatin remodelling complex leads to enrichment of ventral telencephalon progenitors. Specifically, mutating the BAF subunit ARID1B affects the fate transition of progenitors to oligodendrocyte and interneuron precursor cells, a phenotype that we confirmed in patient-specific induced pluripotent stem cell-derived organoids. Our study paves the way for high-throughput phenotypic characterization of disease susceptibility genes in organoid models with cell state, molecular pathway and gene regulatory network readouts.

MeSH terms

  • Autism Spectrum Disorder* / complications
  • Autism Spectrum Disorder* / genetics
  • Autism Spectrum Disorder* / pathology
  • Autistic Disorder / complications
  • Autistic Disorder / genetics
  • Autistic Disorder / pathology
  • Brain* / cytology
  • Brain* / metabolism
  • CRISPR-Associated Protein 9 / metabolism
  • CRISPR-Cas Systems
  • Cell Lineage / genetics
  • Chromatin / genetics
  • Developmental Disabilities* / complications
  • Developmental Disabilities* / genetics
  • Developmental Disabilities* / pathology
  • Gene Editing
  • Humans
  • Loss of Function Mutation
  • Mosaicism
  • Neurons / metabolism
  • Neurons / pathology
  • Organoids* / cytology
  • Organoids* / metabolism
  • RNA, Guide, CRISPR-Cas Systems
  • Single-Cell Gene Expression Analysis*
  • Transcription, Genetic


  • ARID1B protein, human
  • Chromatin
  • CRISPR-Associated Protein 9
  • RNA, Guide, CRISPR-Cas Systems
  • SMARCA2 protein, human
  • SMARCA4 protein, human