Complete Disruption of Autism-Susceptibility Genes by Gene Editing Predominantly Reduces Functional Connectivity of Isogenic Human Neurons.
Complete Disruption of Autism-Susceptibility Genes by Gene Editing Predominantly Reduces Functional Connectivity of Isogenic Human Neurons.
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DOI：
10.1016/j.stemcr.2018.10.003
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发表时间：
2018-11-13
影响因子：
5.9
通讯作者：
Scherer SW
中科院分区：
文献类型：
--
作者：
Deneault E;White SH;Rodrigues DC;Ross PJ;Faheem M;Zaslavsky K;Wang Z;Alexandrova R;Pellecchia G;Wei W;Piekna A;Kaur G;Howe JL;Kwan V;Thiruvahindrapuram B;Walker S;Lionel AC;Pasceri P;Merico D;Yuen RKC;Singh KK;Ellis J;Scherer SW
自闭症谱系障碍（ASD）在表型和基因上具有异质性。我们提出了一种CRISPR基因编辑策略，即插入一个蛋白质标签和提前终止位点，为10个与ASD相关的基因（AFF2/FMR2、ANOS1、ASTN2、ATRX、CACNA1C、CHD8、DLGAP2、KCNQ2、SCN2A、TENM1）的功能研究创建一个诱导多能干细胞（iPSC）敲除资源。神经生成素2（NGN2）引导的iPSC诱导能够产生兴奋性神经元，并且无法检测到突变蛋白。RNA测序揭示了几个神经元网络的汇聚。通过膜片钳和多电极阵列方法，不同突变所测量到的电生理缺陷各不相同。然而，它们最终都导致突触活动的持续减少，包括AFF2/FMR2 -、ASTN2 -、ATRX -、KCNQ2 -和SCN2A - 缺失神经元中自发兴奋性突触后电流频率的降低。尽管ASD易感基因属于不同的基因本体，但同基因干细胞资源能够揭示出共同的功能表型，比如功能连接性降低。
用于10个ASD风险基因功能研究的iPSC敲除资源
与神经元和突触相关的常见转录网络的破坏
在功能多样的ASD风险基因中普遍观察到的突触活动减少
在本文中，谢勒及其同事提出了一种用于10个与自闭症谱系障碍相关基因功能研究的人类诱导多能干细胞（iPSC）敲除资源。他们还表明，这些属于不同功能类别的基因中的一些，可能是CRISPR同基因iPSC衍生的谷氨酸能神经元中共同表型的基础。
Autism spectrum disorder (ASD) is phenotypically and genetically heterogeneous. We present a CRISPR gene editing strategy to insert a protein tag and premature termination sites creating an induced pluripotent stem cell (iPSC) knockout resource for functional studies of ten ASD-relevant genes (AFF2/FMR2, ANOS1, ASTN2, ATRX, CACNA1C, CHD8, DLGAP2, KCNQ2, SCN2A, TENM1). Neurogenin 2 (NGN2)-directed induction of iPSCs allowed production of excitatory neurons, and mutant proteins were not detectable. RNA sequencing revealed convergence of several neuronal networks. Using both patch-clamp and multi-electrode array approaches, the electrophysiological deficits measured were distinct for different mutations. However, they culminated in a consistent reduction in synaptic activity, including reduced spontaneous excitatory postsynaptic current frequencies in AFF2/FMR2-, ASTN2-, ATRX-, KCNQ2-, and SCN2A-null neurons. Despite ASD susceptibility genes belonging to different gene ontologies, isogenic stem cell resources can reveal common functional phenotypes, such as reduced functional connectivity. iPSC knockout resource for functional studies of ten ASD-risk genes Disruption of common transcriptional networks associated with neurons and synapses Reduced synaptic activity commonly observed for functionally diverse ASD-risk genes In this article, Scherer and colleagues present a human induced pluripotent stem cell (iPSC) knockout resource for functional studies of ten genes associated with autism spectrum disorder. They also show that some of these genes, pertaining to diverse functional categories, can underlie common phenotypes in CRISPR-isogenic iPSC-derived glutamatergic neurons.
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DOI：
10.1016/s1474-4422(15)00044-7
发表时间：
2015-11
期刊：
The Lancet. Neurology
影响因子：
--
作者：
Geschwind DH;State MW
通讯作者：
State MW
DOI：
10.1016/j.mcn.2015.12.002
发表时间：
2016-06
期刊：
Molecular and cellular neurosciences
影响因子：
--
作者：
Habela CW;Song H;Ming GL
通讯作者：
Ming GL
影响因子：
3.7
作者：
Brett, Maggie;McPherson, John;Tan, Ene-Choo
通讯作者：
Tan, Ene-Choo
影响因子：
64.5
作者：
Law, Martin J.;Lower, Karen M.;Gibbons, Richard J.
通讯作者：
Gibbons, Richard J.
影响因子：
6.2
作者：
Abrahams BS;Arking DE;Campbell DB;Mefford HC;Morrow EM;Weiss LA;Menashe I;Wadkins T;Banerjee-Basu S;Packer A
通讯作者：
Packer A