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Merck

SCR531

STEMCCA Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit

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UNSPSC Code:
12161503
NACRES:
NA.81
eCl@ss:
32161000

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description

Induced Pluripotent Stem Cells
Mouse Embryonic Stem Cells

technique(s)

cell culture | stem cell: suitable

Quality Level

100

Analysis Note

Tested to confirm the generation of iPS cells from p3 mouse embryonic fibroblasts and p6 human foreskin fibroblasts. Cre-mediated excision of the reprogramming transgenes has only been validated in the mouse system. Other cell types have not been tested and thus similar results can not be guaranteed.

Disclaimer

This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges {HCompany} to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The STEMCCA Cre-Excisable Constitutive Polycistronic (OKSM) Lentivirus Kit contains high titer polycistronic (OKSM) lentivirus and Polybrene transfection reagent that have been validated for the generation of mouse and human induced pluripotent stem cells from mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs), respectively. The Cre-excisable STEMCCA vector, comprised of the transcription factors Oct-4, Klf4, SOX-2, and c-Myc (OKSM) separated by the self-cleaving 2A peptide and IRES sequences, includes flanking LoxP sites that enable Cre-mediated excision of the vector following reprogramming (Sommer CA, 2009).

The use of a single polycistronic lentiviral vector significantly improves reprogramming efficiencies and reduces the number of viral integrations. Furthermore, the ability to remove the reprogramming vector has been shown to improve the developmental potential of iPS cells and significantly increase their capacity to undergo directed differentiation in vitro (Sommer CA, 2010).

Other Notes

1. EF1α-STEMCCA-LoxP (OKSM) Lentivirus: (Part number CS204439) Three (3) vials containing 15 µL of high titer lentivirus. 2. Polybrene Transfection Reagent: (Part number TR-1003-50UL) One (1) vial containing 50 µL of 10 mg/mL stock.

Preparation Note

Lentivirus is stable for at least 6 months when stored at -80°C. After first thaw, place immediately on ice and store in working aliquots to avoid further freeze thaws. Avoid freeze thaws as this will result in a decrease in the virus titer. Polybrene transfection reagent is stable for at least 1 year when stored at -20°C.

Legal Information

Storage Class

10 - Combustible liquids

wgk

WGK 2

Certificates of Analysis (COA)

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Efficient generation of transgene-free human and mouse iPS cells using a cell-permeant Tat-Cre protein
STEMCCA Lentivirus Reprogramming Kits

Millipore’s new STEMCCA lentivirus reprogramming kits make it easier than ever to generate induced pluripotent stem (iPS) cells. Unlike traditional iPS generation which requires simultaneous co-infection by four separate expression vectors, the STEMCCA kits use a single polycistronic lentiviral vector to improve efficiency and reduce the number of viral integrations. The STEMCCA vector is comprised of the transcription factors Oct-4, Klf4, SOX-2, and c-Myc (OKSM), separated by the self-cleaving 2A peptide and IRES sequences 1,2. It is also available with flanking LoxP sites incorporated for Cre-mediated excision of the exogenous reprogramming transgenes. STEMCCA Vector Advantages: (1) Efficient: uses a single vector with four transcription factors rather than co-transducing four separate expression vectors (2) Minimizes viral integrations: single vector reduces the risks of insertional mutagenesis and viral reactivation and (3) Excisable: Cre/LoxP-regulated version enables removal of reprogramming transgenes.

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Global Trade Item Number

SKUGTIN
SCR53104053252014260

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