Targeted release of transcription factors for human cell reprogramming by ZEBRA cell-penetrating peptide.
Transcription factors (TFs) are key actors of the control of gene expression and consequently of every major process within cells, ranging from cell fate determination, cell cycle control and response to environment. Their ectopic expression has proven high potential in reprogramming cells for regenerative medicine; ontogenesis studies and cell based modelling. Direct delivery of proteins could represent an alternative to current reprogramming methods using gene transfer but still needs technological improvements. Herein, we set-up an efficient cellular penetration of recombinant TFs fused to the minimal transduction domain (MD) from the ZEBRA protein. We show that ZEBRA MD-fused TFs applied on primary human fibroblasts and cord blood CD34+ hematopoietic stem cells route through the cytoplasm to the nucleus. The delivery of Oct4, Sox2 and Nanog by MD leads to the activation of mRNA transcripts from genes regulated by these TFs. Moreover, the expression of genes involved in the pluripotency network but not directly bound by these TFs, is also induced. Overall, the repeated application of MD-Oct4, MD-Sox2, MD-Nanog TFs and the post-transcriptional regulator RNA-binding protein MD-Lin28a, triggers the rejuvenation of human fibroblasts and CD34+ cells. This study provides powerful tools for cell fate reprogramming without genetic interferences.
['Caulier|Benjamin|B|', 'Berthoin|Lionel|L|', 'Coradin|Helène|H|', 'Garban|Frédéric|F|', 'Dagher|Marie Claire|MC|', 'Polack|Benoît|B|', 'Toussaint|Bertrand|B|', 'Lenormand|Jean Luc|JL|', 'Laurin|David|D|']
D000818:Animals D057846:Cell-Penetrating Peptides D002478:Cells, Cultured D065150:Cellular Reprogramming D016503:Drug Delivery Systems D005347:Fibroblasts D006412:Hematopoietic Stem Cells D006801:Humans D000071317:Nanog Homeobox Protein D050814:Octamer Transcription Factor-3 D055748:SOXB1 Transcription Factors D014157:Transcription Factors
Transcription factors Reprogramming Cell-penetrating peptide DAPI: 4′,6-diamidino-2-phénylindole (PubChem CID: 2 954) IPTG: Isopropyl-beta-d-thiogalactopyranoside (PubChem CID: 656894) Induced pluripotent stem cells Protein delivery Transdifferentiation Valproic acid (PubChem CID: 3121)