美国哈佛大学Paola Arlotta等研究人员合作利用体内Perturb-Seq技术，揭示出与自闭症风险基因相关的神经元和神经胶质异常。2020年11月27日，国际知名学术期刊《科学》发表了这一成果。

研究人员应用了一种可扩展的遗传筛选方法，叫做体内Perturb-Seq，在功能上评估了35个自闭症谱系障碍/神经发育延迟（ASD/ND）从头丧失功能的风险基因。使用CRISPR-Cas9，研究人员在子宫内发育小鼠大脑内的这些风险基因中引入了移码突变，然后对产后大脑中受扰动的细胞进行单细胞RNA测序。

研究人员从神经元和神经胶质细胞分类中鉴定了细胞类型特异性和进化保守的基因模块。反复的基因模块和细胞类型在整个扰动中都受到影响，这代表了跨越ASD/ND风险基因集的关键细胞效应。体内Perturb-Seq能够研究多样化的突变如何影响发育中生物的细胞类型和状态。

据介绍，通过人类基因研究确定的疾病风险基因和基因座的数量远远超过了系统研究其功能的能力。

附：英文原文

Title: In vivo Perturb-Seq reveals neuronal and glial abnormalities associated with autism risk genes

Author: Xin Jin, Sean K. Simmons, Amy Guo, Ashwin S. Shetty, Michelle Ko, Lan Nguyen, Vahbiz Jokhi, Elise Robinson, Paul Oyler, Nathan Curry, Giulio Deangeli, Simona Lodato, Joshua Z. Levin, Aviv Regev, Feng Zhang, Paola Arlotta

Issue&Volume: 2020/11/27

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.

DOI: 10.1126/science.aaz6063

Source: https://science.sciencemag.org/content/370/6520/eaaz6063