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(NAS Colloquium) Genetic Engineering of Viruses and Viral Vectors (1996) / Chapter Skim
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Alphavirus-based expression vectors: Strategies and applications
Pages 11371-11377

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From page 11371...
... replacement of the structural genes to produce self-replicating RNA "replicons" that can be packaged into infectious particles using defective helper RNAs or packaging cell lines. In addition, incorporation of heterologous ligands or receptors into the virion envelope may eventually allow targeting of engineered alphavirus RNAs to specific cell types.
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From page 11372...
... Translated regions of alphavirus genomic and subgenomic RNAs are shown as boxes with the nonstructural proteins and structural proteins (STRUCTURAL) indicated as open and lightly shaded boxes, respectively.
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From page 11373...
... In addition to packaging of alphavirus RNA replicons by cotransfection with DHRNAs, continuous packaging cell lines have been developed that express a DHRNA under the control of a nuclear promoter (I.F. and S.S., unpublished results)
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From page 11374...
... Using double subgenomic RNA promoter constructs, electroporation and puromycin selection can be used to quickly establish cell populations or clonal cell lines expressing heterologous RNAs and proteins (Fig.
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From page 11375...
... These full-length random insertion libraries are now being used to identify permissive insertion sites for other peptides and larger functional domains that are compatible with recovery of infectious virus. In the case of the Rift Valley fever virus epitope library, replacement of the 4D4-encoding oligonucleotide in the full-length random insertion library with another oligonucleotide can be accomplished in a single step and was used to identify a cluster of sites in the E3 protein permissive for insertion of an 81-residue heterologous peptide (S.
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From page 11376...
... Adaptive mutations are then cloned and mapped by making chimeras with the parental replicon. Modified dsSIN vectors, which include the adaptive mutations and the PAC gene, allow rapid production of BHK cell lines expressing heterologous genes without deleterious effects on the host cell.
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From page 11377...
... colloquium Paper: Frolov et al. Li, G., Barbieri, M
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Key Terms

  • structural proteins
  • rna replicons
  • unpublished results
  • capsid protein
  • cell lines

  • alphavirus rna
  • bhk cells
  • cell death
  • subgenomic rnas
  • rna replication

  • alphavirus rnas
  • mrna translation
  • dssin recombinants
  • replication machinery
  • cell types

  • host mrna
  • packaging machinery
  • infectious particles
  • double subgenomic
  • vertebrate cells

  • engineered alphavirus
  • mammalian cells
  • adaptive mutations
  • noncytopathic gene
  • random insertion

  • viral glycoproteins
  • valley fever
  • transfected cells
  • rna transcripts
  • rna promoter


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