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Safe Production of High-Titer Retrovirus

Safety: Because there is minimal sequence overlap between the packaging vectors and most MMLV retroviral vectors, the probability of generating replication-competent retrovirus (RCR) by homologous recombination is greatly reduced.
  • Compatibility: Compatible selectable markers allow generation of stable producer lines.
  • Versatility: Depending on the range of cell types to be infected, there are several envelope proteins to choose from.

    • Traditionally, stable packaging lines and some vectors for transient virus production were constructed by preserving all or most of the MMLV genomic DNA and deleting the packaging site so the gag-pol and env-expressing genome could not be packaged itself. However, a problem inherent with these systems involves the high degree of homology between the psi-deleted packaging vector and the retroviral expression vector, which allows a high degree of RCR to be generated.1 Thus, these vectors pose a safety problem when vectors containing toxic inserts are pseudotyped with envelope proteins that permit infection of human cells. In the packaging vectors described, the only MMLV-derived sequences are the gag-pol and envelope proteins themselves (except for the VSV-G protein, which is derived from the vesicular stomatatitis virus).

    Host Range of Envelope Proteins (1,5)

    Target Cell

    Ecotropic

    Amphotropic

    10A1

    VSV-G

    Mouse

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