mRNA Reprogramming Factors Set: hOKSML

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000067_figure_1
Morphological timeline for mRNA iPS Colony Formation. Day 1 of chronological morphology changes during the emergence of a primary iPS colony generated using Stemgent’s mRNA Reprogramming Factor Set on Parkinson’s disease patient dermal fibroblasts.
000067 Figure 2
Morphological timeline for mRNA iPS Colony Formation. Day 16 of chronological morphology changes during the emergence of a primary iPS colony generated using Stemgent’s mRNA Reprogramming Factor Set on Parkinson’s disease patient dermal fibroblasts.
000067 Figure 3
Morphological timeline for mRNA iPS Colony Formation. Day 20 of chronological morphology changes during the emergence of a primary iPS colony generated using Stemgent’s mRNA Reprogramming Factor Set on Parkinson’s disease patient dermal fibroblasts.
000067_figure_4
Pluripotent human iPS cell colonies generated with the mRNA Reprogramming System. Phase contrast image of an mRNA-derived iPS cell colony. Immunohistochemistry image of same colony stained for the pluripotency markers. (Nanog/Tra-1-60)
000067 Figure 5
Pluripotent human iPS cell colonies generated with the mRNA Reprogramming System. Phase contrast image of an mRNA-derived iPS cell colony. Immunohistochemistry image of same colony stained for the pluripotency markers. (Oct4/Tra-1-81)
000067_figure_6
Pluripotent human iPS cell colonies generated with the mRNA Reprogramming System. Phase contrast image of an mRNA-derived iPS cell colony. Immunohistochemistry image of same colony stained for the pluripotency markers. (Rex1/SSEA4)
000067_figure7
Relative endogenous mRNA expression level comparison for Nanog and Oct4. qRT-PCR calculations and data normalization against HuES9 p62 human ES positive control cell line.

Product Overview

Stemgent mRNA Reprogramming Factors Set: hOKSML contains mRNA encoding Oct4, Klf4, Sox2, c-Myc, Lin-28, and nuclear GFP (nGFP). These in vitro transcribed mRNAs incorporate both pseudouridine and 5-methylcytidine-modified nucleotides to minimize the cellular interferon response to single stranded RNA.1,2 Repeated transfection of mRNA encoding Oct4, Klf4, Sox2, c-Myc and Lin-28 has been shown to reprogram human fibroblasts.2 nGFP mRNA is included for use as a positive control for RNA transfection during the cellular reprogramming process.

Advantages

  • Faster reprogramming, colonies emerge as early as 16 days
  • Stoichiometric control of individual reprogramming factors
  • Eliminates need for backend screening of viral systems
  • Validated protocol using normal and diseased cells

Product Specifications

Kit / Set Components

2 vials - Stemgent Oct4 mRNA, Human (Cat. No. 05-0014), 20 μg
1 vial - Stemgent Klf4 mRNA, Human (Cat. No. 05-0015), 20 μg
1 vial - Stemgent Sox2, Human (Cat. No. 05-0016), 20 μg
1 vial - Stemgent c-Myc mRNA, Human (Cat. No. 05-0018), 20 μg
1 vial - Stemgent Lin-28 mRNA, Human (Cat. No. 05-0017), 20 μg
1 vial - Stemgent nGFP mRNA (Cat. No. 05-0019), 20 μg

Formulation

100 ng/μl in RNase-free TE buffer, pH 7.0

Storage and Stability

Store at or below -70°C. Stable for a minimum of 3 months from date of receipt when stored as directed. Wear gloves when handling this product. Use RNase-free tubes and barrier pipette tips. Thaw on ice. Keep on ice until the sample is removed. Avoid multiple freezing and thawing cycles (<5). Aliquot to experimental quantities.

Quality Control

Proper protein expression and localization have been confirmed by immunocytochemistry for reprogramming factors and by fluorescence microscopy for nGFP.

Recommended Usage

mRNAs should be transfected using a protocol specifically optimized for the cell type used in the experiment.

Related Products 

  • Stemgent® Oct4 mRNA, human (Cat.No.05-0014 )
  • Stemgent® Klf4 mRNA, Human (Cat.No.05-0015)
  • Stemgent® Sox2 mRNA, Human (Cat.No.05-0016)
  • Stemgent® c-Myc mRNA, Human (Cat.No.05-0018)
  • Stemgent® Lin-28 mRNA, Human (Cat.No.05-0017)
  • Stemgent® nGFP mRNA (Cat.No.05-0019)
  • Pluriton™ mRNA Reprogramming Medium (Cat.No. 00-0070)
  • Stemfect™ RNA Transfection Kit (Cat.No.00-0069)
  • Stemolecule™ Y27632 (Cat.No.04-0012)
  • Stemfactor™ Fibroblast Growth Factor-basic (Human Recombinant) (Cat.No.03-0002)

Documents

Limited Use License MIT

Limited Use License iPS Academia Japan


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Stemgent Comments and Reviews (1)

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Gang Wang It is working well and I already say Bye-Bye to virus
Oct 1, 2011

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