A Novel LHX6 Reporter Cell Line for Tracking Human iPSC-Derived Cortical Interneurons.
A Novel LHX6 Reporter Cell Line for Tracking Human iPSC-Derived Cortical Interneurons.
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作者：
Cruz-Santos M;Cardo LF;Li M
GABA能中间神经元控制大脑中的神经回路和网络活动。皮质中间神经元的功能障碍，特别是那些来自内侧神经节隆起，有助于神经系统疾病的状态。多能干细胞衍生的中间神经元为理解神经精神疾病的病因学提供了有力的工具，并且有可能用作神经系统疾病（如癫痫）的细胞治疗药物。虽然可以在体外容易地诱导大量的中间神经元祖细胞，但是确定的中间神经元亚型的产生仍然是低效的。在hPSC中使用CRISPR/Cas9辅助的同源重组，我们分别将mEmerald和mCherry荧光蛋白的编码序列插入LHX 6的下游，LHX 6是内侧神经节隆起（MGE）衍生的皮质中间神经元所需的基因和标记物。在LHX 6-mEmerald和LHX 6-mCherry hPSC向MGE命运分化后，两种报告基因在hPSC的LHX 6 + MGE衍生物中表现出受限的表达。此外，报告基因的表达对中间神经元诱导信号的变化有反应。因此，LHX 6-报告细胞系代表了鉴定控制人中间神经元发育的分子和设计更好的中间神经元分化方案以及研究与中间神经元病相关的风险基因的有价值的工具。
GABAergic interneurons control the neural circuitry and network activity in the brain. The dysfunction of cortical interneurons, especially those derived from the medial ganglionic eminence, contributes to neurological disease states. Pluripotent stem cell-derived interneurons provide a powerful tool for understanding the etiology of neuropsychiatric disorders, as well as having the potential to be used as medicine in cell therapy for neurological conditions such as epilepsy. Although large numbers of interneuron progenitors can be readily induced in vitro, the generation of defined interneuron subtypes remains inefficient. Using CRISPR/Cas9-assisted homologous recombination in hPSCs, we inserted the coding sequence of mEmerald and mCherry fluorescence protein, respectively, downstream that of the LHX6, a gene required for, and a marker of medial ganglionic eminence (MGE)-derived cortical interneurons. Upon differentiation of the LHX6-mEmerald and LHX6-mCherry hPSCs towards the MGE fate, both reporters exhibited restricted expression in LHX6+ MGE derivatives of hPSCs. Moreover, the reporter expression responded to changes of interneuron inductive cues. Thus, the LHX6-reporter lines represent a valuable tool to identify molecules controlling human interneuron development and design better interneuron differentiation protocols as well as for studying risk genes associated with interneuronopathies.
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影响因子：
23.9
作者：
Colasante, Gaia;Lignani, Gabriele;Broccoli, Vania
通讯作者：
Broccoli, Vania
DOI：
10.1126/science.aar6821
发表时间：
2018-04-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mi D;Li Z;Lim L;Li M;Moissidis M;Yang Y;Gao T;Hu TX;Pratt T;Price DJ;Sestan N;Marín O
通讯作者：
Marín O
影响因子：
16.2
作者：
Nóbrega-Pereira S;Kessaris N;Du T;Kimura S;Anderson SA;Marín O
通讯作者：
Marín O
影响因子：
23.9
作者：
Maroof, Asif M.;Keros, Sotirios;Tyson, Jennifer A.;Ying, Shui-Wang;Ganat, Yosif M.;Merkle, Florian T.;Liu, Becky;Goulburn, Adam;Stanley, Edouard G.;Elefanty, Andrew G.;Widmer, Hans Ruedi;Eggan, Kevin;Goldstein, Peter A.;Anderson, Stewart A.;Studer, Lorenz
通讯作者：
Studer, Lorenz
DOI：
10.1111/j.1460-9568.2012.08156.x
发表时间：
2012-06
期刊：
The European journal of neuroscience
影响因子：
--
作者：
Lewis DA
通讯作者：
Lewis DA