Epi5™ Episomal iPSC Reprogramming Kit

Catalog number: A15960

Invitrogen™  Related applications: Stem Cell Engineering & Reprogramming

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Description

The Epi5™ Episomal iPSC Reprogramming Kit provides an easy-to-use, highly efficient set of 5 episomal vectors designed by Dr. Okita in the laboratory of Professor Yamanaka at the Center for iPS Cell Research and Application (CiRA), Kyoto University. This system produces transgene-free, virus-free human induced pluripotent stem cells (iPSCs) with efficiencies in the range of 0.04% to 0.3%, depending upon the cell type being reprogrammed.

Features of the Epi5™ Episomal iPSC Reprogramming Kit include:

• Easy to use and highly efficient—no need for small molecules during reprogramming and the added p53 suppression provides enhanced iPSC generation (1)
• Usable for all stages of your iPSC research—transgene-free and viral-free reprogramming allows use from basic to pre-clinical research
• Flexibility in media systems—can be used in either feeder-free or feeder-based media systems

Create Transgene and Virus-free iPSCs
Episomal vectors are a well-described system for producing transgene-free, virus-free iPSCs, providing a source of iPSCs for all stages of your pluripotent stem cell research. Other reprogramming methods, such as lentivirus, contain transgenes that can integrate into the host genome, potentially disrupting the genome or causing unpredictable results.

As oriP/EBNA1 vectors, these episomal vectors contain 5 reprogramming factors (Oct4, Sox2, Lin28, Klf4, and L-Myc) and replicate extra-chromosomally only once per cell cycle. At this replication rate, the episomes are lost at a rate of approximately 5% per cell generation. This system shows enhanced iPSC generation through p53 suppression, and the inclusion of L-Myc has been shown to be more potent and specific then c-Myc during human iPSC generation (1).

Easy-to-Use System
This 2-vial kit provides the optimal system for episomal vector reprogramming and eliminates the need for use of small molecules in the reprogramming medium. The episomal vectors are introduced into the cell by electroporation. We recommend use of the Neon®Transfection System for electroporation, allowing flexibility in the volume and thus number of cells needed for reprogramming.

Flexibility in Selection of Media Systems
The Epi5™ Episomal iPSC Reprogramming Kit can be used with multiple cell types and media systems. Protocols for fibroblasts and CD34+ blood cells are available in either feeder or feeder-free based reprogramming.

1. Okita K, Matsumara Y, Sato Y, et al. A more efficient method to generate integration free human iPS cells. Nature Methods 8, 409-412, 2011.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: Cag
Key Function: Episomal Expression, Multi-Gene Expression, Induced Pluripotent Stem Cell Generation
Protein Tag or Fusion: None
Reporter: None
Delivery Method: Transfection
Cloning Method: None
Selection Agent (Eukaryotic): None
Vector Type: EBNA OriP
Product Size: 1 kit
Replication Origin: pUC Origin

Contents & storage

2 tubes. Store at -68 to -85°C.

Documents

Manuals & protocols

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