Nuclear Reprogramming with Hindbrain Patterning Genes by Nanochannel Electroporation

Abstract

Directly inducing a specific cell fate requires not only the knowledge of which transcriptional regulatory networks orchestrate one's desired cellular phenotype, but also the ability to exogenously introduce these proteins into cells. As has been elegantly demonstrated, nuclear reprogramming methodologies typically require small collections of transcription factors to change a cell's epigenetic framework. To generate desired cell types, expression larger collections of patterning genes are needed to generate specific neuronal fates. However, the ability to directly control the dosage of reprogramming factors that each individual cell would need has not been achieved. We have overcome this problem by implementing nanochannel electroporation for neuronal reprogramming. With NEP precise amounts of reprogramming agents are delivered electrophoretically through the nanochennel, with agent dosing being modulated by electrophertic pulse duration and total number of pulses. We utilized the technology to induce specific neuronal cell types of the hindbrain lineage.