Generation of Human Neurons by microRNA-Mediated Direct Conversion of Dermal Fibroblasts
Generation of Human Neurons by microRNA-Mediated Direct Conversion of Dermal Fibroblasts
复制标题
DOI：
10.1007/978-1-0716-1084-8_6
复制
发表时间：
2021-01-01
期刊：
影响因子：
--
通讯作者：
Yoo, Andrew S.
中科院分区：
文献类型：
--
作者：
Church, Victoria A.;Cates, Kitra;Yoo, Andrew S.
MicroRNAs(MiRNAs)、miR-9/9*和miR-124(miR-9/9*-124)在人成纤维细胞中异位表达时，通过消除成纤维细胞的身份并引起泛神经元状态，显示出命运重编程活性。与诱导的多能干细胞来源的神经元不同，miRNA诱导的神经元(Mins)通过直接命运转换保留了起始成纤维细胞的生物学年龄，从而提供了一个基于人类神经元的平台，用于研究老年神经元固有的细胞特性，并使用患者来源的细胞模拟成人起病的神经退行性疾病。此外，神经元亚型特异性转录因子的表达与miR-9/9*-124一起引导MIN走向不同的神经元命运，这一特征对于影响特定神经元亚型的疾病建模至关重要。在这里，我们描述了基于miR-9/9*-124的神经元重编程协议，用于生成几种与疾病相关的神经元亚型：纹状体中棘神经元、皮质神经元和脊髓运动神经元。
MicroRNAs (miRNAs), miR-9/9*, and miR-124 (miR-9/9*-124) display fate-reprogramming activities when ectopically expressed in human fibroblasts by erasing the fibroblast identity and evoking a pan-neuronal state. In contrast to induced pluripotent stem cell-derived neurons, miRNA-induced neurons (miNs) retain the biological age of the starting fibroblasts through direct fate conversion and thus provide a human neuron-based platform to study cellular properties inherent in aged neurons and model adult-onset neurodegenerative disorders using patient-derived cells. Furthermore, expression of neuronal subtype-specific transcription factors in conjunction with miR-9/9*-124 guides the miNs to distinct neuronal fates, a feature critical for modeling disorders that affect specific neuronal subtypes. Here, we describe the miR-9/9*-124-based neuronal reprogramming protocols for the generation of several disease-relevant neuronal subtypes: striatal medium spiny neurons, cortical neurons, and spinal cord motor neurons.