Puro.Cre empty vector
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||17408||Plasmid sent as bacteria in agar stab||1||$65|
Virus (1 mL at titer ≥ 1x10⁶ TU/mL)
and Plasmid. More Information
This material is available to academics and nonprofits only.
Backbone manufacturerJacks Lab
- Backbone size (bp) 9200
Vector typeMammalian Expression, Lentiviral, Cre/Lox
Growth in Bacteria
- Cloning method Restriction Enzyme
- 5′ sequencing primer n/a
- 3′ sequencing primer MSCV rev (Common Sequencing Primers)
Puro.Cre was initially generated by amplification of PuroR from MSCV.Puro (Clontech) with PuroF (5’-CGCTGCGCCGAATTCATGACCGAGTACAAGCCCACG-3’) and PuroR (5’-TAAGCGGCCGCTCGAGTCAGGCACCGGGCTTGCGGGTC-3’), digestion with EcoRI and NotI and cloning into UbC.Luciferase.PGK.Cre to generate Puro.Cre.
For lentiviral packaging, you can use pCMV-delta-8.2 (Addgene #8455) and pCMV-VSV-G (Addgene #8454).
Information for Lentiviral Prep (Catalog # 17408-LV) ( Back to top )
Ready-to-use Lentiviral Prep particles produced from Puro.Cre empty vector (#17408). In addition to the viral particles, you will also receive purified Puro.Cre empty vector plasmid DNA.Lentiviral particles carrying CRE recombinase and puromycin resistance.
- Volume 1 mL
- Titer ≥1x10⁶ TU/mL
- Pricing $220 USD for preparation of 1 mL virus + $30 USD for plasmid.
- Storage Store at -80℃. Thaw just before use and keep on ice.
- Shipment Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide
Viral Quality Control
- Cell survival assay relative to standard curve: Lenti-X 293T cells were serially transduced with 17408-LV or a control virus of known titer. 48h after transduction, cells were selected with puromycin, expanded for 2 days and resistant cells quantified via a dye-based cell viability assay. A standard curve was generated from the serial dilutions of the control virus and used to determine the titer of 17408-LV.
- PCR confirmation of insert: PCR was carried out with primers targeting CRE and WPRE. The PCR product was visualized on an agarose gel for size confirmation.
Forward Primer: CMV-F TTTACGGCGCTAAGGATGAC
Reverse Primer: WPRE-R CATAGCGTAAAAGGAGCAACA
- Confirmation of Cre activity: Expression of Cre following transduction with 17408-LVC was confirmed using a Cre reporter assay.
- Toxicity: Please note that that Cre expression is toxic in some cell lines due to cryptic loxP sites in the genome. If you are unsure how your cell line of interest tolerates Cre expression, consider transducing with a range MOIs or transducing an array of cell lines.
Visit our viral production page for more information.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:Puro.Cre empty vector was a gift from Tyler Jacks (Addgene plasmid # 17408)
For your References section:Suppression of non-small cell lung tumor development by the let-7 microRNA family. Kumar MS, Erkeland SJ, Pester RE, Chen CY, Ebert MS, Sharp PA, Jacks T. Proc Natl Acad Sci U S A. 2008 Feb 28. ():. 10.1073/pnas.0712321105 PubMed 18308936