title: Cellular manipulation of human embryonic stem cells by TAT-PDX1 protein transduction.
abstract: Human embryonic stem cells (hESCs) are an in vitro model system for the study of human early development and a potential source for cell-based therapies. An efficient strategy for cellular manipulation of hESCs may be highly valuable for the analysis of gene function involved in human embryogenesis and the development of cell-based therapies via induced differentiation into particular cell types. However, plasmid transfection of hESCs has low efficiency and viral transduction may not be the method of choice for cell-based therapies due to genome integration. To overcome these limitations, we applied protein transduction technology that can transfer proteins into cells via direct penetration across the lipid bilayer. Here, we show that the FITC dye fused to the TAT protein transduction domain (PTD) was efficiently transferred into hESCs. In addition, the PDX1 transcription factor, which plays a central role in pancreatic development, was transferred into hESCs as a fusion form of TAT PTD. The transduced TAT-PDX1 activated its downstream target genes and induced insulin protein production in hESCs. These results demonstrate that protein transduction could be used in the cellular manipulation of hESCs and would provide a significant breakthrough for basic and therapeutic research in hESCs.
authors.affiliation: Institute of Reproductive Medicine and Population, Medical Research Center, Korea.
authors.forename: Young Do
authors.initials: YD
authors.lastname: Kwon
authors.forename: Sun Kyung
authors.initials: SK
authors.lastname: Oh
authors.forename: Hee Sun
authors.initials: HS
authors.lastname: Kim
authors.forename: Seung-Yup
authors.initials: SY
authors.lastname: Ku
authors.forename: Seok Hyun
authors.initials: SH
authors.lastname: Kim
authors.forename: Young Min
authors.initials: YM
authors.lastname: Choi
authors.forename: Shin Yong
authors.initials: SY
authors.lastname: Moon
mesh_terms: D000818:Animals; D002460:Cell Line; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D015696:Gene Products, tat / Q000235:genetics* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology; D014161:Transduction, Genetic
chemical_list: D015696:Gene Products, tat; D018398:Homeodomain Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D016650:Fluorescein-5-isothiocyanate