Neuroscience and Mental Health Research Institute, School of Medicine, Cardiff University, Cardiff CF24 4HQ, UK.
School of Biosciences, Cardiff University, Cardiff CF10 3AX, UK.
Cells. 2022 Mar 1;11(5):853. doi: 10.3390/cells11050853.
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 , 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.
GABA 能中间神经元控制大脑的神经回路和网络活动。皮质中间神经元的功能障碍，特别是源自内侧神经节隆起的中间神经元的功能障碍，导致神经病变状态。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提供了一个强大的工具，并且有可能在细胞治疗中用作治疗神经疾病的药物，如癫痫。尽管可以在体外很容易地诱导大量中间神经元前体，但定义的中间神经元亚型的产生仍然效率低下。我们使用 CRISPR/Cas9 辅助同源重组在 hPSC 中，分别在 基因的下游插入 mEmerald 和 mCherry 荧光蛋白的编码序列， 基因是 所必需的，也是内侧神经节隆起（MGE）衍生的皮质中间神经元的标志物。当 LHX6-mEmerald 和 LHX6-mCherry hPSC 向 MGE 命运分化时，两个报告器都在 hPSC 的 LHX6 MGE 衍生物中表现出受限表达。此外，报告器表达响应于中间神经元诱导线索的变化。因此，LHX6 报告基因系代表了一种有价值的工具，可以识别控制人类中间神经元发育的分子，并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。
