In vivo Perturb-Seq reveals neuronal and glial abnormalities associated with autism risk genes.
In vivo Perturb-Seq reveals neuronal and glial abnormalities associated with autism risk genes.
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DOI：
10.1126/science.aaz6063
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发表时间：
2020-11-27
期刊：
影响因子：
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通讯作者：
Arlotta P
中科院分区：
文献类型：
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作者：
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;Arlotta P
通过人类遗传学研究确定的疾病风险基因和位点的数量远远超过了我们系统研究其功能的能力。我们应用了一种可扩展的遗传筛查方法，体内Perturb-Seq，对35个自闭症谱系障碍/神经发育迟缓（ASD/ND）从头功能丧失风险基因进行功能评估。使用CRISPR-Cas9，我们在子宫内发育的小鼠大脑中的这些风险基因中引入移码突变，然后对出生后大脑中受干扰的细胞进行单细胞RNA测序。我们确定了细胞类型特异性和进化保守的基因模块从神经元和神经胶质细胞类。复发基因模块和细胞类型在这组扰动中受到影响，代表了ASD/ND风险基因组的关键细胞效应。体内Perturb-Seq使我们能够研究不同的突变如何影响发育中生物体的细胞类型和状态。子宫内的Perturb-Seq遗传筛查揭示了大脑发育过程中一组精神疾病风险基因扰动影响的复发性和细胞类型特异性基因模块。
The number of disease risk genes and loci identified through human genetic studies far outstrips our 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. An in utero Perturb-Seq genetic screen reveals recurrent and cell type-specific gene modules affected across a panel of psychiatric disorder risk gene perturbations during brain development.