Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production

Retrovirus vectors can be made in the absence of helper virus by using retrovirus packaging cell lines. Helper-free virus is critical for a variety of gene transfer studies. The most useful packaging cell lines contain helper virus DNA from which the signal required for packaging of the viral RNA genome into virions has been deleted. However, we showed that the ability to package virus is conferred at very low frequency to cells infected with virus from these packaging cell lines, presumably by low-frequency transmission of the deleted virus genome. In addition, these packaging cell lines can interact with some retroviral vectors to yield replication-competent virus. We constructed packaging cell lines containing helper virus DNA that had several alterations in addition to deletion of the packaging signal. The new packaging cells retained the useful features of previously available lines but did not yield helper virus after introduction of any of the vectors tested, and transfer of the packaging function was not detected.

[1]  A. Miller,et al.  Retroviral transfer of genes into canine hemopoietic progenitor cells in culture: a model for human gene therapy. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[2]  A. Miller,et al.  Retrovirus-mediated transfer and expression of drug resistance genes in human haematopoietic progenitor cells , 1986, Nature.

[3]  A. Miller,et al.  Factors involved in production of helper virus-free retrovirus vectors , 1986, Somatic cell and molecular genetics.

[4]  P. Kantoff,et al.  Gene expression in mice after high efficiency retroviral-mediated gene transfer. , 1985, Science.

[5]  G. Keller,et al.  Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors , 1985, Nature.

[6]  J. Dick,et al.  Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice , 1985, Cell.

[7]  D. Baltimore,et al.  Varying the position of a retrovirus packaging sequence results in the encapsidation of both unspliced and spliced RNAs , 1985, Journal of virology.

[8]  I. Verma,et al.  Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene , 1985, Molecular and cellular biology.

[9]  R. Mulligan,et al.  High-efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Sorge,et al.  Amphotropic retrovirus vector system for human cell gene transfer , 1984, Molecular and cellular biology.

[11]  I. Verma,et al.  Two base changes restore infectivity to a noninfectious molecular clone of Moloney murine leukemia virus (pMLV-1) , 1984, Journal of virology.

[12]  H. Temin,et al.  Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors , 1983, Molecular and cellular biology.

[13]  H. Temin,et al.  The terminal nucleotides of retrovirus DNA are required for integration but not virus production , 1983, Nature.

[14]  I. Verma,et al.  A transmissible retrovirus expressing human hypoxanthine phosphoribosyltransferase (HPRT): gene transfer into cells obtained from humans deficient in HPRT. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Bohlen,et al.  Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyl transferase. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Baltimore,et al.  Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus , 1983, Cell.

[17]  E. Scolnick,et al.  Construction and isolation of a transmissible retrovirus containing the src gene of Harvey murine sarcoma virus and the thymidine kinase gene of herpes simplex virus type 1 , 1981, Journal of virology.

[18]  M. Pearson,et al.  Enhancing the efficiency of DNA-mediated gene transfer in mammalian cells , 1981, Somatic cell genetics.

[19]  M. Linial,et al.  Avian oncovirus mutant (SE21Q1b) deficient in genomic RNA: characterization of a deletion in the provirus , 1980, Journal of virology.

[20]  K. Quade.  Transformation of mammalian cells by avian myelocytomatosis virus and avian erythroblastosis virus. , 1979, Virology.

[21]  P. Kourilsky,et al.  Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in Escherichia coli K-12. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Hartley,et al.  Naturally occurring murine leukemia viruses in wild mice: characterization of a new "amphotropic" class , 1976, Journal of virology.

[23]  M. Gardner,et al.  Amphotropic host range of naturally occuring wild mouse leukemia viruses , 1976, Journal of virology.

[24]  W. Russell,et al.  A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses , 1975, Nature.

[25]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[26]  J. Hartley,et al.  Plaque assay techniques for murine leukemia viruses. , 1970, Virology.