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Title: Generation of non-viral, transgene-free hepatocyte like cells with piggyBac transposon
Authors: Katayama, Hokahiro
Yasuchika, Kentaro
Miyauchi, Yuya
Kojima, Hidenobu
Yamaoka, Ryoya
Kawai, Takayuki
Yukie Yoshitoshi, Elena
Ogiso, Satoshi
Kita, Sadahiko
Yasuda, Katsutaro
Sasaki, Naoya
Fukumitsu, Ken
Komori, Junji
Ishii, Takamichi  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0002-7461-9653 (unconfirmed)
Uemoto, Shinji  kyouindb  KAKEN_id
Author's alias: 安近, 健太郎
Issue Date: 15-Mar-2017
Publisher: Springer Nature
Journal title: Scientific Reports
Volume: 7
Thesis number: 44498
Abstract: Somatic cells can be reprogrammed to induced hepatocyte-like cells (iHeps) by overexpressing certain defined factors in direct reprogramming techniques. Of the various methods to deliver genes into cells, typically used genome-integrating viral vectors are associated with integration-related adverse events such as mutagenesis, whereas non-integrating viral vectors have low efficiency, making viral vectors unsuitable for clinical application. Therefore, we focused on developing a transposon system to establish a non-viral reprogramming method. Transposons are unique DNA elements that can be integrated into and removed from chromosomes. PiggyBac, a type of transposon, has high transduction efficiency and cargo capacity, and the integrated transgene can be precisely excised in the presence of transposase. This feature enables the piggyBac vector to achieve efficient transgene expression and a transgene-free state, thus making it a promising method for cell reprogramming. Here, we attempted to utilize the piggyBac transposon system to generate iHeps by integrating a transgene consisting of Hnf4a and Foxa3, and successfully obtained functional iHeps. We then demonstrated removal of the transgene to obtain transgene-free iHeps, which still maintained hepatocyte functions. This non-viral, transgene-free reprogramming method using the piggyBac vector may facilitate clinical applications of iHeps in upcoming cell therapy.
Rights: © The Author(s) 2017. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
URI: http://hdl.handle.net/2433/219318
DOI(Published Version): 10.1038/srep44498
PubMed ID: 28295042
Appears in Collections:Journal Articles

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