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Versatile Reporter Vectors for Monitoring Viral Transduction



Determine retroviral transduction efficiency and gene expression in virtually any target cell line

Brenda Rogers Katherine Felts Peter Vaillancourt
Stratagene

We describe three new ViraPort retroviral reporter vectors: pFB-hrGFP Y, pFB-Neo-LacZ ,, and pFB-Luc , which can be used to determine transduction efficiency and gene expression in virtually any target cell line. To assay for expression, use simple fluorescence microscopy or FACS with pFB-hrGFP, carry out a b-galactosidase-activity assay or in situ staining with pFB-Neo-LacZ, or perform a luciferase activity assay with pFB-Luc. These control vectors are available as plasmid DNA as well as transduction-ready, replication-incompetent, VSV-G pseudotyped high-titer retroviral supernatants. The vectors are particularly useful as controls when using the ViraPort retroviral gene expression systems.

We recently introduced a set of human and rodent ViraPort retroviral cDNA expression libraries inserted into the MMLV-based high-titer retroviral vector pFB1,2. These complex cDNA libraries, part of our growing collection of ViraPort retroviral premade libraries, are easily and efficiently introduced into virtually any mitotic cell type for screening based on gene function. The insert capacity of pFB, our MMLV-based replication-defective pFB vector, is approximately 8.0 kb because there is no extraneous sequence between the LTRs. We excluded selectable markers or readily detectable reporter genes from the ViraPort retroviral gene expression systems to ensure that the largest cDNAs possible could be inserted.

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