Vector map of the Precision LentiORF RFP control
For gene-specific constructs, the RFP insert would be replaced by the ORF of interest
Precision LentiORFs are fully-sequenced human open reading frames (ORFs) that have been cloned into a lentiviral expression vector. Ideal for use in primary or difficult to transfect cells, Precision LentiORFs can be used to create stably expressing cell lines, investigate overexpression phenotypes, or perform RNAi rescue experiments.
Features of the Precision LentiORF backbone include a CMV promoter for robust protein expression, a GFP reporter to visually track protein expression, and blasticidin resistance for selection of stable cell lines. Precision LentiORF clones are available with or without native stop codons, and secondary tags can be added to the MultiTag cloning site located downstream of the coding sequence.
- Lentiviral ORF constructs require no subcloning for mammalian expression
- Fully sequenced coding sequences
- GFP reporter is transported to the nucleusfor easy visualization of expression
- Clones are available with or without native stop codons
- Libraries are available as arrayed glycerol stocks
- Individual Clones– Glycerol Stock
- Pre-defined gene families- Arrayed glycerol stocks (includes with and without native stop clones)
- Complete genome-scale collection- Arrayed glycerol stocks(includes with and without native stop clones)
Potential applications of Precision LentiORFs include gene overexpression experiments, RNAi rescue (see the RNAi Rescue using Precision LentiORF technical note), and expression of human proteins in mammalian cell lines. Primary and non-dividing cells can be readily transduced with ready-to-use lentiviral particles transfected with Precision LentiORF constructs directly into your cell line. For generating your own viral particles, Trans-Lentiviral ORF packaging kits are also available.
Clones are provided as a live culture in a 2mL tube. Each tube contains LB medium supplemented with 8% glycerol and the appropriate antibiotic. Within 2 to 3 business days of receiving your order we will ship your clone at room temperature via express delivery. Store the stock clone at –80°C.
Orders of 50 clones or greater and stock plates will be provided in 96-well microtiter plates. These will ship on dry-ice via overnight delivery and should be stored at –80°C immediately upon receipt. Please contact our customer service department for a quote and estimated shipping time on bulk orders.
Due to the inherent complexity of high-titer lentiviral particle production, we estimate a 6 to 8 week turnaround time for these orders.
The Precision LentiORF expression vectors are not compatible with third generation packaging systems. We recommend the Trans-Lentiviral Packaging System for use with these vectors.
Dharmacon is a distributor of multiple gene expression clone collections (cDNAs and ORFs). These clone collections were generated by groups outside of Dharmacon and thus the quality of the collections is largely dependent upon what Dharmacon received from these groups. Specific clone information and plate coordinates were provided to Dharmacon by the suppliers of these clone collections. Dharmacon has not sequence verified each individual clone from these collections. These collections and individual clones are distributed "as is" with no additional product validation or guarantees. Dharmacon has established quality procedures to ensure that individual clones are picked from the identified well in a plate, grown on the correct antibiotic, and are free of phage contamination. Due to the quality of the information provided to Dharmacon, the clone you receive might not match the expected clone. If this occurs, please contact Technical Support.
|Shipping Condition||Dry Ice|
|Storage Conditions||-80 C|
|Stability at Recommended Storage Conditions||At least 12 months|
Visually track Precision LentiORF expression
Nuclear localized TurboGFP expressed from cells transduced with Precision LentiORF viral particles.
Cell cycle phenotypes with ORF expressing PLK1.
Immunocytochemistry of HEK293T cells showing cellular morphology of cytoplasm (Anti-tubulin - red) and nucleus (DAPI - blue).
pLOC vector map
pLOC vector map
Optimize Precision LentiORF expresson level by adjusting MOI
Western blot analysis shows that the level of expression can be optimized by controlling the number of viral particles per cell.
E. M. Briso, J. Guinea-Viniegra, L. Bakiri, Z. Rogon, P. Petzelbauer, R. Eils, R. Wolf, M. Rincón, P. Angel, and E. F. Wagner, Inflammation-mediated skin tumorigenesis induced by epidermal c-Fos. Genes & Dev.27:1959-1973 (Sep 2013).
A. V. Shinde, R. K. Motiani, X. Zhang, I. F. Abdullaev, A. P. Adam, J. C. González-Cobos, W. Zhang, K. Matrougui, P. A. Vincent, and M. Trebak, STIM1 Controls Endothelial Barrier Function Independently of Orai1 and Ca2+ Entry. Sci. Signal.6:ra18 (Mar 2013).
H. Fayyad-Kazan, R. Rouas, M. Merimi, N. El Zein, P. Lewalle, F. Jebbawi, M. Mourtada, H. Badran, M. Ezzeddine, B. Salaun, P. Romero, A. Burny, P. Martiat, and B. Badran, Valproate Treatment of Human Cord Blood CD4-positive Effector T Cells Confers on Them the Molecular Profile (MicroRNA Signature and FOXP3 Expression) of Natural Regulatory CD4-positive Cells through Inhibition of Histone Deacetylase. J. Biol. Chem.285:20481-20491 (Jul 2010).