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Construction of Retro viral Packaging Cell Lines

  • Olivier Danos
Part of the Methods in Molecular Biology book series (MIMB, volume 8)

Abstract

Most of the time, retrovirus vectors retain only cis-acting sequences from the original viral genome. These sequences allow the recombinant structure to be transcribed (LTR promoter/enhancer) and the RNA to be processed (splicing and polyadenylation signals), packaged into a virion particle (packaging sequences), and replicated by the reverse transcriptase (tRNA binding site, R region, and polypurine track). The other viral functions have to be provided in trans for the assembly of recombinant viral particles to take place. This can be simply achieved by using a replication-competent helper virus, leading to the production of a mixed population. Nevertheless, helper-free stocks are desirable for most applications, since
  1. 1.

    The high frequency of recombination in a mixed virus stock is likely to lead to the appearance of recombinants with unknown structure and activity. These new chimeras, either spread by the helper virus or replication-competent themselves, create a potential safety problem.

     
  2. 2.

    Cell lineage analysis using retroviral marking can only be performed and interpreted in a helper-free context

     
  3. 3.

    In vivo gene transfer experiments can be jeopardized by disease (s) associated with helper-virus infection (1).

     

Keywords

Selectable Marker Helper Virus Packaging Cell Line Reverse Transcriptase Assay Indicator Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© The Humana Press Inc., Clifton, NJ 1991

Authors and Affiliations

  • Olivier Danos
    • 1
  1. 1.Institut PasteurParisFrance

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