Revolutionary Technology for Efficient Derivation of Human iPSCs with Messenger R

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DESCRIPTION (provided by applicant): The goal of the proposed project is to develop a commercial kit that will allow drug addiction researchers with ongoing projects using cell-based models to be able to successfully and conveniently produce human induced pluripotent stem cells (iPSCs). Addiction to drugs destroys lives of substance abusers and their families, creating enormous social problems and exerting cost on the nation's health care system. Molecular mechanisms that contribute to the initiation and establishment of addiction are largely unknown, partially due to the lack of neural cell types from the patients. The technologies that made possible of human iPSCs opened the doors to new areas of direct interrogation of the pathophysiology of addiction by using patient-specific cell sources. Currently, the NIDA is funding projects that rely on iPS technologies to: 1) characterize dopaminergic neurotransmission function of neurons derived from iPSCs with genomic aberrations, such as chromosomal polymorphism regions containing dopamine transporter (DAT) and vesicular monoamine transporter (VMAT2) genes; 2) identify genetic variations that can be functionally correlated to addiction in patient cell-derived neurons via iPSCs; 3) make dopamine neurons from patient iPSCs and expose them to methamphetamine and withdrawal in order to analyze the electrophysiological properties of patient-specific neurons versus controls, etc. Even though it is considered a technology with great potential in studies of addiction or other diseases, reprogramming of adult human cells to iPSCs is still an inefficient, tedious and costly process to most researchers. The currently prevailing methods of reprogramming have additional problems such as alteration of genomes or introduction of viruses that can complicate the interpretation of results from the cell-based assays. This project aims to develop a turn-key tool kit that will enable many biomedical researchers to generate iPSCs not only with 10 to 100 folds higher efficiency than other methods, in less than 2 weeks, but also through a feeder-free, xeno-free and footprint-free procedure.