( High transduction efficiency a...)High-Titer ,Retroviral Vectors for Gene Delivery

High transduction efficiency and predictable copy number

High-Titer Retroviral Vectors for Gene Delivery

Katherine Felts John C. Bauer Peter Vaillancourt
Stratagene

The high-titer retroviral vectors pFB and pFB-Neo can be used with any Moloney Murine Leukemia Virus (MMLV)-based packaging cell line. Virus produced with these vectors can be used to infect a wide range of cell types including human cells, and because MMLV-based virions are labile and readily inactivated by ethanol, ultraviolet light, and human complement, they are relatively safe and can be used in a P2 tissue culture facility. In transient virus production experiments, using a 293 cell-based producer line, we have achieved titers in excess of 10 ⁸ colony-forming units per ml (cfu/ml) of supernatant for pFB, and greater than 10⁷ cfu/ml for pFB-Neo. Protein expression from the viral promoter in infected cells was comparable to or higher than that for the vector pLXSN, another commercially available MMLV vector. The vector pFB has been deleted for all viral coding sequence and contains no exogenous sequence other than that for the multiple cloning site (MCS). As has been shown for other such minimal MMLV-based vectors, pFB is capable of insertions of up to 8 kb without appreciable loss of titer.¹

The development of high-titer retroviral vectors that are capable of infecting a wide variety of cell types has had a tremendous impact on fields for which highly efficient gene delivery is essential.² When tissue culture cells are infected with virus harboring a gene of interest, such experiments commonly result in transduction efficiencies of greater than 90%. An additional advantage of retroviral vectors is that the copy number of integrated provirus can be easily controlled
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