Non-Integrating iPSC Generation

Since 2006 when Takahashi et al. described the ability to reprogram adult cells to Induced Pluripotent Stem (iPS) cells through the overexpression of transcription factors, iPS has been a widely studied field due to its potential for treating disease.  However, till now the best method for iPSC generation has been lentiviral transmission of transcription factors.  Since 2009 Allele Biotechnology has offered various viral tools for iPS induction: retroviral vectors and lentiviral particles of all iPS factors, a 4-in-1 lentivirus with OSKM in one virus, p53 shRNA to boost transfection efficiency, and a Cre-LoxP system to remove the viral genome after reprograming.  When using viruses, even those flanked by loxP sites, there is still a rearrangement of the cell’s genome; and a chance that viral inserts remain. A footprint of the virus remains and for use in a clinical setting, this footprint is not an acceptable outcome.

In 2010, Warren et al. reported a novel, footprint-free method to reprogram human fibroblasts into iPSCs using messenger RNA.  In our latest publication, "Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA" which appears in Scientific Reports, Dr. Warren and the team of scientists at Allele Bitoechnology present a substantially enhanced mRNA reprogramming protocol.  By harnessing engineered reprograming factors and optimizing various experimental conditions we can now offer a feeder-free, xeno-free protocol that can reprogram human skin cells to pluripotency in under two weeks. 

We are now offering 6F mRNA Premix, which contains mRNA for the 6 reprogramming factors used in the new protocol combined in the appropriate ratios.  The premix is validated and ready for direct use in your iPSC experiments. 

  • Inherently non-mutagenic vector for footprint-free iPSC production
  • Reprogramming of human fibroblasts with 9 days of transfection
  • Eliminates safety concerns and bio-containment issues associated with virus
  • Completely feeder-free and xeno-free reprogramming protocol



Allele Biotechnology / 2012  ripsc-staining-website.jpg


         Immunostaining for cell-surface and nuclear pluripotency markers.  Copyright Allele Biotech 2012.  


NextGen mRNA Based iPSC Derivation

There are three options for harnessing our NextGen mRNA based iPSC reprogramming and differentiation technology, 6F mRNA premix, iPSC generation custom services, and IVT templates for generating synthetic mRNA.

mRNA Premix

iPSC Derivation Service

IVT Templates

6F mRNA Reprogramming Premix contains all the factors necessary for accelerated iPSC reprogramming in one convenient mix.   The mix is funtionally validated and ready for use in your reprogramming experiments.  One vial of premix is sufficient to conduct feeder-free reprogramming on eight wells in a 6-well plate format.  Our next-gen mRNA technology allows for highly efficient iPSC generation in less time then any other current methods.

Custom iPSC generation services are an excellent option for deriving pluripotent cell lines.  Each project is tailored to fit your needs.  We start with your target cell line and generate iPSCs using our own next-gen mRNA platform. Once colonies are obtained you choose your deliverables and the desired validation.

In vitro trancription (IVT) templates are ready-to-use linear DNA templates for making synthetic mRNA. We currently offer IVT templates for iPSC generation and neural differentiation, and offer new template construction as a custom service.

Click here to view the 6F mRNA Reprogramming Premix product page Click here to view our custom iPSC service page

Click here to view our list of IVT Templates

Click here to view our differentiation factor request page



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Allele Biotechnology / 2012ripsc-time-website.jpg

  Bright-field images taken during the derivation of RNA-iPSCs (RiPS) from human fibroblasts showing colony emergence over a 9 day time frame.  Copyright Allele Biotechnology, 2012. 



Kazutoshi Takahashi and Shinya Yamanaka. (2006)  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors.  Cell doi:10.1016/j.cell.2006.07.024

Luigi Warren, Philip D. Manos, Tim Ahfeldt, Yuin-Han Loh, Hu Li, Frank Lau, Wataru Ebina, Pankaj K. Mandal, Zachary D. Smith, Alexander Meissner, George Q. Daley, Andrew S. Brack, James J. Collins, Chad Cowan, Thorsten M. Schlaeger  and Derrick J. Rossi (2010) Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA. Cell Stem Cell, doi:10.1016/j.stem.2010.08.012