title: Transcription factor-based modulation of neural stem cell differentiation using direct protein transduction.
abstract: In contrast to conventional gene transfer strategies, the direct introduction of recombinant proteins into cells bypasses the risk of insertional mutagenesis and offers an alternative to genetic intervention. Here, we explore whether protein transduction of the gliogenic transcription factor Nkx2.2 can be used to promote oligodendroglial differentiation of mouse embryonic stem cell (ESC)-derived neural stem cells (NSC). To that end, a recombinant cell-permeant form of Nkx2.2 protein was generated. Exposure of ESC-derived NSC to the recombinant protein and initiation of differentiation resulted in a two-fold increase in the number of oligodendrocytes. Furthermore, Nkx2.2-transduced cells exhibited a more mature oligodendroglial phenotype. Comparative viral gene transfer studies showed that the biological effect of Nkx2.2 protein transduction is comparable to that obtained by lentiviral transduction. The results of this proof-of-concept study depict direct intracellular delivery of transcription factors as alternative modality to control lineage differentiation in NSC cultures without genetic modification.
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE and BRAIN Center, University of Bonn, Bonn, Germany.
authors.forename: Kristin
authors.initials: K
authors.lastname: Stock
authors.forename: Lars
authors.initials: L
authors.lastname: Nolden
authors.forename: Frank
authors.initials: F
authors.lastname: Edenhofer
authors.forename: Tamara
authors.initials: T
authors.lastname: Quandel
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D048430:Cell Shape; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D005822:Genetic Vectors; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D009687:Nuclear Proteins; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D029961:Zebrafish Proteins
chemical_list: D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; C000715631:NKX2-2 protein, human; C000715627:Nkx2-2 protein, mouse; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; C000715630:nkx2.2b protein, zebrafish