Description
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
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.
Research Category
Stem Cell Research
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.
