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Product Specification Sheet
Product Name
Stemgent® Dox Inducible Reprogramming Lentivirus Set: mouse OKSM (conc)
Description
The Stemgent® Dox Inducible Reprogramming Lentivirus Set: mouse OKSM (conc)
contains doxycycline (dox)-inducible lentiviruses capable of expressing one of four
mouse transcription factors (Oct4, Sox2, Klf4 or c-Myc) under the control of the doxinducible tetO operator when transduced into mammalian cells. Each unique virus is
provided as a VSV-G pseudotyped and concentrated virus stock capable of infecting
both dividing and non-dividing cells. The expression of these four transcription
factors, along with the reverse tetracycline transcriptional activator (rtTA), has been
shown to reprogram a variety of mouse cell types including fibroblasts, B cells, T
cells, and neural precursor cells to an embryonic stem (ES) cell-like state known as
the induced pluripotent stem (iPS) cell1. Proper isolation and subsequent injection of
these iPS cells into blastocysts can generate mice that carry all four inducible viruses,
which can be reactivated upon the addition of dox2. The Dox Inducible GFPLentivirus is included as a transduction control.
Catalog Number
ST000014
Set Contents
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100 µl Dox Inducible Reprogramming Lentivirus Mouse Oct4 (conc)
(Cat. No. ST070003)
100 µl Dox Inducible Reprogramming Lentivirus Mouse Klf4 (conc)
(Cat. No. ST070005)
100 µl Dox Inducible Reprogramming Lentivirus Mouse Sox2 (conc)
(Cat. No. ST070002)
100 µl Dox Inducible Reprogramming Lentivirus Mouse c-Myc (conc)
(Cat. No. ST070004)
4 x 50 µl lentivirus rtTA (Cat. No. ST070048)
100 µl Dox Inducible Lentivirus GFP (Cat. No. ST070049)
Storage and Stability
Store at -70°C for up to 3 months. Keep frozen until use. Avoid freeze thaw cycles.
Quality Control
Viral integration copy number was quantified by transduction of mammalian cells
followed by qPCR of the genomic DNA for PBS/psi. Transcription factor protein
expression was verified by immunocytochemistry analysis of transduced mammalian
cells. Reprogramming was verified by transduction of mouse embryonic fibroblasts,
generation of iPS cell colonies, and subsequent colony characterization by
immunocytochemistry. Tested to be negative for Mycoplasma sp. by PCR and
microbial contamination by a sterility test.
Safety Precautions
The greatest safety risk associated with viral delivery systems stems from the
potential generation of recombinant viruses that are capable of autonomous
replication during the packaging process. The Stemgent Reprogramming Lentivirus
platform eliminates these hazards by combining a disabled viral genome with a
unique manufacturing process. Also, gene functions that facilitate the enclosing of
the silencing sequence in a viral capsid (e.g., Gag, Pol, Env) are distributed into
multiple helper plasmids (which do not contain significant regions of homology)
during packaging. This tactic further minimizes the probability of recombination
events that might otherwise generate viruses capable of autonomous replication.
With these safety measures in place, the Stemgent Reprogramming Lentivirus
particles can be employed in standard Biosafety Level 2 tissue culture facilities and
3050 Spruce Street
Saint Louis, Missouri 63103 USA
Telephone 800-325-5832 - (314) 771-5765
Fax (314) 286-7828
email: [email protected]
sigma-aldrich.com
1
Product Specification Sheet
should be treated with the same level of caution as with any other potentially
infectious agent.
References
1.
2.
Brambrink, T., Foreman, R., Welstead, G.G., Lengner, C.J., Wernig, M., Suh, H.,
and Jaenisch, R. (2008) Sequential expression of pluripotency markers during
direct reprogramming of mouse somatic cells. Cell Stem Cell 2, 151-159.
Wernig, M., Lengner, C.J., Hanna, J., Lodato, M.A., Steine, E., Foreman, R.,
Staerk, J., Markoulaki, S., and Jaenisch, R. (2008) A drug-inducible transgenic
system for direct reprogramming of multiple somatic cell types. Nat Biotechnol
26, 916-924.
Restricted Use
For restrictions related to the use of this product please refer to the accompanying
Restricted Use License.
Notice to Purchaser
Purchaser represents and warrants that it will use the Stemgent Dox Inducible
Reprogramming Lentivirus Set: mouse OKSM (conc) purely for research purposes,
not for diagnostic use, not for resale, and not for use in humans or veterinary
applications. Stemgent will not be held responsible for patent infringement or other
violations that may occur with the use of our products.
Stemgent Reprogramming Lentivirus products are sold through Sigma-Aldrich Inc.
Sigma-Aldrich Inc. warrants that its products conform to the information contained
in this and in other Sigma-Aldrich publications. Purchaser must determine the
suitability of the product(s) for their particular use. Additional terms and conditions
may apply. Please see the reverse side of the invoice or packing slip.
3050 Spruce Street
Saint Louis, Missouri 63103 USA
Telephone 800-325-5832 - (314) 771-5765
Fax (314) 286-7828
email: [email protected]
sigma-aldrich.com
2