Synthetically modified mRNA for efficient and fast human iPS cell generation and direct transdifferentiation to myoblasts

Synthetic mRNA transfection enables efficient and controlled gene expression in human cells, without genome integrations. Further, modifications to the mRNA and transfection protocol now allow for repeated transfection and long-term gene expression of an otherwise short-lived mRNA expression. This i...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2016-05, Vol.473 (3), p.743-751
Main Authors: Preskey, David, Allison, Thomas F., Jones, Mark, Mamchaoui, Kamel, Unger, Christian
Format: Article
Language:English
Subjects:
Cell Lineage
Cell Transdifferentiation
Cellular Reprogramming
Fibroblast
fibroblasts
Fibroblasts - cytology
gene expression
Gene Expression Profiling
Gene Expression Regulation
Human iPS
Humans
Induced Pluripotent Stem Cells - cytology
interferons
Interferons - metabolism
In vitro
In vitro transcription
In vitro
In vitro transcription
messenger RNA
Myoblast
myoblasts
Myoblasts - cytology
MyoD Protein - metabolism
MYOD1
nucleosides
Nucleosides - metabolism
Reprogramming
RNA, Messenger - metabolism
Synthetic mRNA
Transdifferentiation
Transfection
transgenes
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Summary:Synthetic mRNA transfection enables efficient and controlled gene expression in human cells, without genome integrations. Further, modifications to the mRNA and transfection protocol now allow for repeated transfection and long-term gene expression of an otherwise short-lived mRNA expression. This is mainly achieved through introducing modified nucleosides and interferon suppression. In this study we provide an overview and details of the advanced synthesis and modifications of mRNA originally developed for reprogramming. This mRNA allows for very efficient transfection of fibroblasts enabling the generation of high quality human iPS cells with a six-factor mRNA cocktail in 9 days. Furthermore, we synthesised and transfected modified MYOD1 mRNA to transdifferentiate human fibroblasts into myoblast-like cells without a transgene footprint. This efficient and integration-free mRNA technology opens the door for safe and controlled gene expression to reverse or redirect cell fate.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2015.09.102