CRISPR/Cas9-mediated heterozygous knockout of the autism gene CHD8 and characterization of its transcriptional networks in neurodevelopment.
Ping Wang,Mingyan Lin,Erika Pedrosa,Anastasia Hrabovsky,Zheng Zhang,Wenjun Guo,Herbert M. Lachman,Deyou Zheng +7 more
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TLDR
Transcriptome profiling revealed that CHD8 hemizygosity (CHD8+/−) affected the expression of several thousands of genes in neural progenitors and early differentiating neurons, and seven of the twelve genes associated with human brain volume or head size by genome-wide association studies were dysregulated in CHD 8+/+ neural progensitors or neurons.Abstract:
Disruptive mutation in the CHD8 gene is one of the top genetic risk factors in autism spectrum disorders (ASDs). Previous analyses of genome-wide CHD8 occupancy and reduced expression of CHD8 by shRNA knockdown in committed neural cells showed that CHD8 regulates multiple cell processes critical for neural functions, and its targets are enriched with ASD-associated genes. To further understand the molecular links between CHD8 functions and ASD, we have applied the CRISPR/Cas9 technology to knockout one copy of CHD8 in induced pluripotent stem cells (iPSCs) to better mimic the loss-of-function status that would exist in the developing human embryo prior to neuronal differentiation. We then carried out transcriptomic and bioinformatic analyses of neural progenitors and neurons derived from the CHD8 mutant iPSCs. Transcriptome profiling revealed that CHD8 hemizygosity (CHD8
+/−) affected the expression of several thousands of genes in neural progenitors and early differentiating neurons. The differentially expressed genes were enriched for functions of neural development, β-catenin/Wnt signaling, extracellular matrix, and skeletal system development. They also exhibited significant overlap with genes previously associated with autism and schizophrenia, as well as the downstream transcriptional targets of multiple genes implicated in autism. Providing important insight into how CHD8 mutations might give rise to macrocephaly, we found that seven of the twelve genes associated with human brain volume or head size by genome-wide association studies (e.g., HGMA2) were dysregulated in CHD8
+/− neural progenitors or neurons. We have established a renewable source of CHD8
+/− iPSC lines that would be valuable for investigating the molecular and cellular functions of CHD8. Transcriptomic profiling showed that CHD8 regulates multiple genes implicated in ASD pathogenesis and genes associated with brain volume.read more
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CRISPR-Based Technologies for the Manipulation of Eukaryotic Genomes.
Alexis C. Komor,Alexis C. Komor,Alexis C. Komor,Ahmed H. Badran,Ahmed H. Badran,Ahmed H. Badran,David R. Liu,David R. Liu,David R. Liu +8 more
TL;DR: Recent developments that extend the generality, DNA specificity, product selectivity, and fundamental capabilities of natural CRISPR systems are described.
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Genetic compensation induced by deleterious mutations but not gene knockdowns
TL;DR: In this article, the authors show that egfl7 mutants do not show any obvious phenotypes while animals injected with egfl 7 morpholino (morphants) exhibit severe vascular defects, indicating that the activation of a compensatory network to buffer against deleterious mutations was not observed after translational or transcriptional knockdown.
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Applications of CRISPR technologies in research and beyond
TL;DR: Programmable DNA cleavage using CRISPR–Cas9 enables efficient, site-specific genome engineering in single cells and whole organisms and is being used to expedite crop and livestock breeding, engineer new antimicrobials and control disease- carrying insects with gene drives.
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CRISPR/Cas9-mediated heterozygous knockout of the autism gene CHD8 and characterization of its transcriptional networks in cerebral organoids derived from iPS cells
Ping Wang,Ryan Mokhtari,Erika Pedrosa,Michael Kirschenbaum,Can Bayrak,Deyou Zheng,Herbert M. Lachman +6 more
TL;DR: The findings show that distinct ASD, SZ, and BD candidate genes converge on common molecular targets—an important consideration for developing novel therapeutics in genetically heterogeneous complex traits.
Journal ArticleDOI
Chd8 mediates cortical neurogenesis via transcriptional regulation of cell cycle and Wnt signaling.
Omer Durak,Omer Durak,Fan Gao,Yea Jin Kaeser-Woo,Richard Rueda,Anthony J Martorell,Anthony J Martorell,Alexi Nott,Carol Y. Liu,L Ashley Watson,Li-Huei Tsai,Li-Huei Tsai,Li-Huei Tsai +12 more
TL;DR: It is reported that Chd8 knockdown during cortical development results in defective neural progenitor proliferation and differentiation that ultimately manifests in abnormal neuronal morphology and behaviors in adult mice, and Transcriptome analysis revealed that while Chd 8 stimulates the transcription of cell cycle genes, it also precludes the induction of neural-specific genes by regulating the expression of PRC2 complex components.
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