Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Jun;70(6):3922–3929. doi: 10.1128/jvi.70.6.3922-3929.1996

Gene transfer into mammalian cells by a Rous sarcoma virus-based retroviral vector with the host range of the amphotropic murine leukemia virus.

E V Barsov 1, S H Hughes 1
PMCID: PMC190270  PMID: 8648729

Abstract

We have constructed and characterized a Rous sarcoma virus-based retroviral vector with the host range of the amphotropic murine leukemia virus (MLV). The chimeric retroviral genome was created by replacing the env coding region in the replication-competent retroviral vector RCASBP(A) with the env region from an amphotropic MLV. The recombinant vector RCASBP-M(4070A) forms particles containing MLV Env glycoproteins. The vector replicates efficiently in chicken embryo fibroblasts and is able to transfer genes into mammalian cells. Vector stocks with titers exceeding 10(6) CFU/ml on mammalian cells can be easily prepared by passaging transfected chicken embryo fibroblasts. Since the vector is inherently defective in mammalian cells, it appears to have the safety features required for gene therapy.

Full Text

The Full Text of this article is available as a PDF (583.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Astrin S. M., Buss E. G., Haywards W. S. Endogenous viral genes are non-essential in the chicken. Nature. 1979 Nov 15;282(5736):339–341. doi: 10.1038/282339a0. [DOI] [PubMed] [Google Scholar]
  2. Bates P., Young J. A., Varmus H. E. A receptor for subgroup A Rous sarcoma virus is related to the low density lipoprotein receptor. Cell. 1993 Sep 24;74(6):1043–1051. doi: 10.1016/0092-8674(93)90726-7. [DOI] [PubMed] [Google Scholar]
  3. Berberich S. L., Macias M., Zhang L., Turek L. P., Stoltzfus C. M. Comparison of Rous sarcoma virus RNA processing in chicken and mouse fibroblasts: evidence for double-spliced RNA in nonpermissive mouse cells. J Virol. 1990 Sep;64(9):4313–4320. doi: 10.1128/jvi.64.9.4313-4320.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boerkoel C. F., Federspiel M. J., Salter D. W., Payne W., Crittenden L. B., Kung H. J., Hughes S. H. A new defective retroviral vector system based on the Bryan strain of Rous sarcoma virus. Virology. 1993 Aug;195(2):669–679. doi: 10.1006/viro.1993.1418. [DOI] [PubMed] [Google Scholar]
  5. Buchschacher G. L., Jr, Panganiban A. T. Human immunodeficiency virus vectors for inducible expression of foreign genes. J Virol. 1992 May;66(5):2731–2739. doi: 10.1128/jvi.66.5.2731-2739.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
  7. Cone R. D., Mulligan R. C. High-efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6349–6353. doi: 10.1073/pnas.81.20.6349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cosset F. L., Legras C., Chebloune Y., Savatier P., Thoraval P., Thomas J. L., Samarut J., Nigon V. M., Verdier G. A new avian leukosis virus-based packaging cell line that uses two separate transcomplementing helper genomes. J Virol. 1990 Mar;64(3):1070–1078. doi: 10.1128/jvi.64.3.1070-1078.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cosset F. L., Ronfort C., Molina R. M., Flamant F., Drynda A., Benchaibi M., Valsesia S., Nigon V. M., Verdier G. Packaging cells for avian leukosis virus-based vectors with various host ranges. J Virol. 1992 Sep;66(9):5671–5676. doi: 10.1128/jvi.66.9.5671-5676.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Deng W. P., Nickoloff J. A. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal Biochem. 1992 Jan;200(1):81–88. doi: 10.1016/0003-2697(92)90280-k. [DOI] [PubMed] [Google Scholar]
  11. Donahue R. E., Kessler S. W., Bodine D., McDonagh K., Dunbar C., Goodman S., Agricola B., Byrne E., Raffeld M., Moen R. Helper virus induced T cell lymphoma in nonhuman primates after retroviral mediated gene transfer. J Exp Med. 1992 Oct 1;176(4):1125–1135. doi: 10.1084/jem.176.4.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dong J., Roth M. G., Hunter E. A chimeric avian retrovirus containing the influenza virus hemagglutinin gene has an expanded host range. J Virol. 1992 Dec;66(12):7374–7382. doi: 10.1128/jvi.66.12.7374-7382.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dubay J. W., Roberts S. J., Hahn B. H., Hunter E. Truncation of the human immunodeficiency virus type 1 transmembrane glycoprotein cytoplasmic domain blocks virus infectivity. J Virol. 1992 Nov;66(11):6616–6625. doi: 10.1128/jvi.66.11.6616-6625.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Federspiel M. J., Bates P., Young J. A., Varmus H. E., Hughes S. H. A system for tissue-specific gene targeting: transgenic mice susceptible to subgroup A avian leukosis virus-based retroviral vectors. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11241–11245. doi: 10.1073/pnas.91.23.11241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Federspiel M. J., Crittenden L. B., Hughes S. H. Expression of avian reticuloendotheliosis virus envelope confers host resistance. Virology. 1989 Nov;173(1):167–177. doi: 10.1016/0042-6822(89)90232-8. [DOI] [PubMed] [Google Scholar]
  16. Gebhardt A., Bosch J. V., Ziemiecki A., Friis R. R. Rous sarcoma virus p19 and gp35 can be chemically crosslinked to high molecular weight complexes. An insight into virus assembly. J Mol Biol. 1984 Apr 5;174(2):297–317. doi: 10.1016/0022-2836(84)90340-1. [DOI] [PubMed] [Google Scholar]
  17. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  18. Granowitz C., Colicelli J., Goff S. P. Analysis of mutations in the envelope gene of Moloney murine leukemia virus: separation of infectivity from superinfection resistance. Virology. 1991 Aug;183(2):545–554. doi: 10.1016/0042-6822(91)90983-i. [DOI] [PubMed] [Google Scholar]
  19. Greenhouse J. J., Petropoulos C. J., Crittenden L. B., Hughes S. H. Helper-independent retrovirus vectors with Rous-associated virus type O long terminal repeats. J Virol. 1988 Dec;62(12):4809–4812. doi: 10.1128/jvi.62.12.4809-4812.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  21. Horton R. M., Cai Z. L., Ho S. N., Pease L. R. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. Biotechniques. 1990 May;8(5):528–535. [PubMed] [Google Scholar]
  22. Hughes S. H., Greenhouse J. J., Petropoulos C. J., Sutrave P. Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors. J Virol. 1987 Oct;61(10):3004–3012. doi: 10.1128/jvi.61.10.3004-3012.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hughes S. H., Kosik E., Fadly A. M., Salter D. W., Crittenden L. B. Design of retroviral vectors for the insertion of foreign deoxyribonucleic acid sequences into the avian germ line. Poult Sci. 1986 Aug;65(8):1459–1467. doi: 10.3382/ps.0651459. [DOI] [PubMed] [Google Scholar]
  24. Hughes S. H., Petropoulos C. J., Federspiel M. J., Sutrave P., Forry-Schaudies S., Bradac J. A. Vectors and genes for improvement of animal strains. J Reprod Fertil Suppl. 1990;41:39–49. [PubMed] [Google Scholar]
  25. Kawai S., Nishizawa M., Shinno-Kohno H., Shiroki K. A variant Schmidt-Ruppin strain of Rous sarcoma virus with increased affinity for mammalian cells. Jpn J Cancer Res. 1989 Dec;80(12):1179–1185. doi: 10.1111/j.1349-7006.1989.tb01652.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Knight J. B., Stinski M. F., Stoltzfus C. M. Avian sarcoma virus RNA synthesis, RNA splicing and virus production in human foreskin fibroblasts: effect of co-infection with human cytomegalovirus. J Gen Virol. 1993 Dec;74(Pt 12):2629–2636. doi: 10.1099/0022-1317-74-12-2629. [DOI] [PubMed] [Google Scholar]
  27. Landau N. R., Page K. A., Littman D. R. Pseudotyping with human T-cell leukemia virus type I broadens the human immunodeficiency virus host range. J Virol. 1991 Jan;65(1):162–169. doi: 10.1128/jvi.65.1.162-169.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miller A. D., Garcia J. V., von Suhr N., Lynch C. M., Wilson C., Eiden M. V. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J Virol. 1991 May;65(5):2220–2224. doi: 10.1128/jvi.65.5.2220-2224.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Miller D. G., Edwards R. H., Miller A. D. Cloning of the cellular receptor for amphotropic murine retroviruses reveals homology to that for gibbon ape leukemia virus. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):78–82. doi: 10.1073/pnas.91.1.78. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nasioulas G., Hughes S. H., Felber B. K., Whitcomb J. M. Production of avian leukosis virus particles in mammalian cells can be mediated by the interaction of the human immunodeficiency virus protein Rev and the Rev-responsive element. Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11940–11944. doi: 10.1073/pnas.92.25.11940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ott D., Friedrich R., Rein A. Sequence analysis of amphotropic and 10A1 murine leukemia viruses: close relationship to mink cell focus-inducing viruses. J Virol. 1990 Feb;64(2):757–766. doi: 10.1128/jvi.64.2.757-766.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ott D., Rein A. Basis for receptor specificity of nonecotropic murine leukemia virus surface glycoprotein gp70SU. J Virol. 1992 Aug;66(8):4632–4638. doi: 10.1128/jvi.66.8.4632-4638.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Otto E., Jones-Trower A., Vanin E. F., Stambaugh K., Mueller S. N., Anderson W. F., McGarrity G. J. Characterization of a replication-competent retrovirus resulting from recombination of packaging and vector sequences. Hum Gene Ther. 1994 May;5(5):567–575. doi: 10.1089/hum.1994.5.5-567. [DOI] [PubMed] [Google Scholar]
  34. Page K. A., Landau N. R., Littman D. R. Construction and use of a human immunodeficiency virus vector for analysis of virus infectivity. J Virol. 1990 Nov;64(11):5270–5276. doi: 10.1128/jvi.64.11.5270-5276.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Perez L. G., Davis G. L., Hunter E. Mutants of the Rous sarcoma virus envelope glycoprotein that lack the transmembrane anchor and cytoplasmic domains: analysis of intracellular transport and assembly into virions. J Virol. 1987 Oct;61(10):2981–2988. doi: 10.1128/jvi.61.10.2981-2988.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Petropoulos C. J., Hughes S. H. Replication-competent retrovirus vectors for the transfer and expression of gene cassettes in avian cells. J Virol. 1991 Jul;65(7):3728–3737. doi: 10.1128/jvi.65.7.3728-3737.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Quintrell N., Hughes S. H., Varmus H. E., Bishop J. M. Structure of viral DNA and RNA in mammalian cells infected with avian sarcoma virus. J Mol Biol. 1980 Nov 15;143(4):363–393. doi: 10.1016/0022-2836(80)90218-1. [DOI] [PubMed] [Google Scholar]
  38. Rein A., Mirro J., Haynes J. G., Ernst S. M., Nagashima K. Function of the cytoplasmic domain of a retroviral transmembrane protein: p15E-p2E cleavage activates the membrane fusion capability of the murine leukemia virus Env protein. J Virol. 1994 Mar;68(3):1773–1781. doi: 10.1128/jvi.68.3.1773-1781.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Salmons B., Moritz-Legrand S., Garcha I., Günzburg W. H. Construction and characterization of a packaging cell line for MMTV-based conditional retroviral vectors. Biochem Biophys Res Commun. 1989 Mar 31;159(3):1191–1198. doi: 10.1016/0006-291x(89)92236-5. [DOI] [PubMed] [Google Scholar]
  40. Salter D. W., Smith E. J., Hughes S. H., Wright S. E., Crittenden L. B. Transgenic chickens: insertion of retroviral genes into the chicken germ line. Virology. 1987 Mar;157(1):236–240. doi: 10.1016/0042-6822(87)90334-5. [DOI] [PubMed] [Google Scholar]
  41. Salter D. W., Smith E. J., Hughes S. H., Wright S. E., Fadly A. M., Witter R. L., Crittenden L. B. Gene insertion into the chicken germ line by retroviruses. Poult Sci. 1986 Aug;65(8):1445–1458. doi: 10.3382/ps.0651445. [DOI] [PubMed] [Google Scholar]
  42. Shimada T., Fujii H., Mitsuya H., Nienhuis A. W. Targeted and highly efficient gene transfer into CD4+ cells by a recombinant human immunodeficiency virus retroviral vector. J Clin Invest. 1991 Sep;88(3):1043–1047. doi: 10.1172/JCI115365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Spector D. H., Wade E., Wright D. A., Koval V., Clark C., Jaquish D., Spector S. A. Human immunodeficiency virus pseudotypes with expanded cellular and species tropism. J Virol. 1990 May;64(5):2298–2308. doi: 10.1128/jvi.64.5.2298-2308.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Valsesia-Wittmann S., Drynda A., Deléage G., Aumailley M., Heard J. M., Danos O., Verdier G., Cosset F. L. Modifications in the binding domain of avian retrovirus envelope protein to redirect the host range of retroviral vectors. J Virol. 1994 Jul;68(7):4609–4619. doi: 10.1128/jvi.68.7.4609-4619.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vanin E. F., Kaloss M., Broscius C., Nienhuis A. W. Characterization of replication-competent retroviruses from nonhuman primates with virus-induced T-cell lymphomas and observations regarding the mechanism of oncogenesis. J Virol. 1994 Jul;68(7):4241–4250. doi: 10.1128/jvi.68.7.4241-4250.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vara J. A., Portela A., Ortín J., Jiménez A. Expression in mammalian cells of a gene from Streptomyces alboniger conferring puromycin resistance. Nucleic Acids Res. 1986 Jun 11;14(11):4617–4624. doi: 10.1093/nar/14.11.4617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vogt V. M., Bruckenstein D. A., Bell A. P. Avian sarcoma virus gag precursor polypeptide is not processed in mammalian cells. J Virol. 1982 Nov;44(2):725–730. doi: 10.1128/jvi.44.2.725-730.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weiss R. A., Boettiger D., Murphy H. M. Pseudotypes of avian sarcoma viruses with the envelope properties of vesicular stomatitis virus. Virology. 1977 Feb;76(2):808–825. doi: 10.1016/0042-6822(77)90261-6. [DOI] [PubMed] [Google Scholar]
  49. Weiss R. A., Wong A. L. Phenotypic mixing between avian and mammalian RNA tumor viruses: I. Envelope pseudotypes of Rous sarcoma virus. Virology. 1977 Feb;76(2):826–834. doi: 10.1016/0042-6822(77)90262-8. [DOI] [PubMed] [Google Scholar]
  50. Whitcomb J. M., Hughes S. H. Retroviral reverse transcription and integration: progress and problems. Annu Rev Cell Biol. 1992;8:275–306. doi: 10.1146/annurev.cb.08.110192.001423. [DOI] [PubMed] [Google Scholar]
  51. Whitcomb J. M., Ortiz-Conde B. A., Hughes S. H. Replication of avian leukosis viruses with mutations at the primer binding site: use of alternative tRNAs as primers. J Virol. 1995 Oct;69(10):6228–6238. doi: 10.1128/jvi.69.10.6228-6238.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Young J. A., Bates P., Willert K., Varmus H. E. Efficient incorporation of human CD4 protein into avian leukosis virus particles. Science. 1990 Dec 7;250(4986):1421–1423. doi: 10.1126/science.2175047. [DOI] [PubMed] [Google Scholar]
  53. Yu X., Yuan X., McLane M. F., Lee T. H., Essex M. Mutations in the cytoplasmic domain of human immunodeficiency virus type 1 transmembrane protein impair the incorporation of Env proteins into mature virions. J Virol. 1993 Jan;67(1):213–221. doi: 10.1128/jvi.67.1.213-221.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. de la Luna S., Soria I., Pulido D., Ortín J., Jiménez A. Efficient transformation of mammalian cells with constructs containing a puromycin-resistance marker. Gene. 1988;62(1):121–126. doi: 10.1016/0378-1119(88)90585-9. [DOI] [PubMed] [Google Scholar]
  55. van Zeijl M., Johann S. V., Closs E., Cunningham J., Eddy R., Shows T. B., O'Hara B. A human amphotropic retrovirus receptor is a second member of the gibbon ape leukemia virus receptor family. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1168–1172. doi: 10.1073/pnas.91.3.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES