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SCR511

Sigma-Aldrich

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

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About This Item

UNSPSC Code:
12352207
eCl@ss:
32161000
NACRES:
NA.81
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manufacturer/tradename

STEMCCA

Quality Level

technique(s)

cell culture | stem cell: suitable

input

sample type: mouse embryonic stem cell(s)
sample type induced pluripotent stem cell(s)

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).

Important note: the STEMCCA lentivirus reprogramming kits are provided in two formats, dependent on whether you are working with a human or mouse system. Human reprogramming experiments require a higher multiplicity of infection (MOI) than mouse experiments. Therefore, the larger, three-vial kits are required for human reprogramming experiments. If you are reprogramming mouse embryo fibroblast cells, you have the option to use either the one- or three-vial kit, depending on how many wells you want to infect. Reprogramming other rodent somatic cells may require more than one vial of lentivirus to perform at optimal MOI.

Application

Research Category
Stem Cell Research
The STEMCCA Cre-Excisable Constitutive Polycistronic Lentivirus Kit contains high titer polycistronic lentivirus & Polybrene transfection reagent that have been validated for the generation of mouse & human induced pluripotent stem cells.

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

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

Other Notes

1. EF1α-STEMCCA-LoxP (OKSM) Lentivirus: (Part number CS204439) One (1) vial 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.

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.

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

监管及禁止进口产品
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Related Content

This guide outlines our comprehensive selection of stem cells, media, supplements, growth factors, cultureware, and tools for stem cell characterization. These proven solutions cover a broad spectrum of stem cell and specialty cell culture areas and are backed by our knowledgeable technical support.

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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