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Cellular manipulation of human embryonic stem cells by TAT-PDX1 protein transduction

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Authors
Kwon, Young Do; Oh, Sun Kyung; Kim, Hee Sun; Ku, Seung-Yup; Kim, Seok Hyun; Choi, Young Min; Moon, Shin Yong
Issue Date
2005-06-21
Publisher
Elsevier
Citation
Mol Ther. 2005 Jul;12(1):28-32.
Keywords
AnimalsCell LineFluorescein-5-isothiocyanate/metabolismGene Products, tat/*genetics/metabolismHomeodomain Proteins/*genetics/metabolismHumansPeptide Fragments/genetics/metabolismRabbitsRecombinant Fusion Proteins/*genetics/metabolismReverse Transcriptase Polymerase Chain Reaction*Stem CellsTrans-Activators/*genetics/metabolismTranscription, Genetic/physiology*Transduction, Genetic
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.
ISSN
1525-0016 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15963917

http://hdl.handle.net/10371/15114
DOI
https://doi.org/10.1016/j.ymthe.2005.03.010
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College of Medicine/School of Medicine (의과대학/대학원)Obstetrics & Gynecology (산부인과전공)Journal Papers (저널논문_산부인과학전공)
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