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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.
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.
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.
Clones are provided as a live culture in a 2 mL 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.
Nuclear localized TurboGFP expressed from cells transduced with Precision LentiORF viral particles.
Immunocytochemistry of HEK293T cells showing cellular morphology of cytoplasm (Anti-tubulin - red) and nucleus (DAPI - blue).
pLOC vector map
Western blot analysis shows that the level of expression can be optimized by controlling the number of viral particles per cell.