Generation of non-viral, transgene-free hepatocyte like cells with piggyBac transposon
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...
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Published in | Scientific reports Vol. 7; no. 1; p. 44498 |
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Main Authors | , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
London
Nature Publishing Group UK
15.03.2017
Nature Publishing Group |
Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/srep44498 |