Nuclear Delivery of Recombinant OCT4 by Chitosan Nanoparticles for Transgene-free Generation of Protein-induced Pluripotent Stem Cells

Overview

Journal Oncotarget

Specialty Oncology

Date 2016 May 18

PMID 27183911

Citations 8

Authors

Salma Tammam

Peter Malak

Daphne Correa

Oliver Rothfuss

Hassan M E Azzazy

Alf Lamprecht

Klaus Schulze-Osthoff

Affiliations

Soon will be listed here.

Abstract

Protein-based reprogramming of somatic cells is a non-genetic approach for the generation of induced pluripotent stem cells (iPSCs), whereby reprogramming factors, such as OCT4, SOX2, KLF4 and c-MYC, are delivered as functional proteins. The technique is considered safer than transgenic methods, but, unfortunately, most protein-based protocols provide very low reprogramming efficiencies. In this study, we developed exemplarily a nanoparticle (NP)-based delivery system for the reprogramming factor OCT4. To this end, we expressed human OCT4 in Sf9 insect cells using a baculoviral expression system. Recombinant OCT4 showed nuclear localization in Sf9 cells indicating proper protein folding. In comparison to soluble OCT4 protein, encapsulation of OCT4 in nuclear-targeted chitosan NPs strongly stabilized its DNA-binding activity even under cell culture conditions. OCT4-loaded NPs enabled cell treatment with high micromolar concentrations of OCT4 and successfully delivered active OCT4 into human fibroblasts. Chitosan NPs therefore provide a promising tool for the generation of transgene-free iPSCs.

Citing Articles

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