Patient-derived or genomically modified human induced pluripotent stem cells (iPSCs) offer the opportunity to study neurodevelopmental and neurodegenerative disorders. Overexpression of certain neurogenic transcription factors (TFs) in iPSCs can induce efficient differentiation into homogeneous populations of the disease-relevant neuronal cell types. Here we provide protocols for genomic manipulations of iPSCs by CRISPR/Cas9. We also introduce two methods, based on lentiviral delivery and the piggyBac transposon system, to stably integrate neurogenic TFs into human iPSCs. Furthermore, we describe the TF-mediated neuronal differentiation and maturation in combination with astrocyte cocultures.
CITATION STYLE
Sauter, E. J., Kutsche, L. K., Klapper, S. D., & Busskamp, V. (2019). Induced neurons for the study of neurodegenerative and neurodevelopmental disorders. In Methods in Molecular Biology (Vol. 1942, pp. 101–121). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9080-1_9
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