1887

Abstract

A method was developed to generate a complex cDNA expression library within an adenovirus type 5 (Ad5)-based vector backbone, termed AdLibrary. Construction of the AdLibrary entailed the conversion of an Ad5 genome-containing cosmid to infectious virus particles. The Ad5 genome was modified by replacing the E1A and E1B genes with a Rous sarcoma virus-driven expression cassette. Conversion was accomplished by liberating the viral genome by restriction enzyme digestion and transfection in HEK 293 cells, which support the growth of E1A/E1B-deficient virus. A test AdLibrary demonstrated the possibility of converting and identifying a marker gene present at a frequency of 1/10 in the cosmid library. To demonstrate the utility of this technology, an AdLibrary was used to isolate a viral gene by its biological function. Virus growth was selected for with an AdLibrary on A549 cells, which do not complement for E1A/E1B function. The AdLibrary was generated with cDNAs derived from HeLa cells productively infected with Ad5. A cDNA corresponding to Ad5 E1A 13S was selected and isolated from the AdLibrary using this strategy. Since multiple genes are assayed simultaneously, this technology should expedite the discovery of genes affecting defined biological activities. This AdLibrary approach provides an opportunity to exploit the efficient gene delivery capabilities of adenovirus vectors for the rapid discovery of gene and protein function.

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2003-12-01
2019-12-11
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References

  1. Berk, A. J. & Sharp, P. A. ( 1977; ). Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell 12, 721–732.[CrossRef]
    [Google Scholar]
  2. Berk, A. J. & Sharp, P. A. ( 1978; ). Structure of the adenovirus 2 early mRNAs. Cell 14, 695–711.[CrossRef]
    [Google Scholar]
  3. Berk, A. J., Lee, F., Harrison, T., Williams, J. & Sharp, P. A. ( 1979; ). Pre-early adenovirus 5 gene product regulates synthesis of early viral messenger RNAs. Cell 17, 935–944.[CrossRef]
    [Google Scholar]
  4. Berkner, K. L. & Sharp, P. A. ( 1983; ). Generation of adenovirus by transfection of plasmids. Nucleic Acids Res 11, 6003–6020.[CrossRef]
    [Google Scholar]
  5. Chartier, C., Degryse, E., Gantzer, M., Dieterle, A., Pavirani, A. & Mehtali, M. ( 1996; ). Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli. J Virol 70, 4805–4810.
    [Google Scholar]
  6. Chow, L. T., Broker, T. R. & Lewis, J. B. ( 1979; ). Complex splicing patterns of RNAs from the early regions of adenovirus-2. J Mol Biol 134, 265–303.[CrossRef]
    [Google Scholar]
  7. Crouzet, J., Naudin, L., Orsini, C. & 11 other authors ( 1997; ). Recombinational construction in Escherichia coli of infectious adenoviral genomes. Proc Natl Acad Sci U S A 94, 1414–1419.[CrossRef]
    [Google Scholar]
  8. Elahi, S. M., Oualikene, W., Naghdi, L., O'Connor-McCourt, M. & Massie, B. ( 2002; ). Adenovirus-based libraries: efficient generation of recombinant adenoviruses by positive selection with the adenovirus protease. Gene Ther 9, 1238–1246.[CrossRef]
    [Google Scholar]
  9. Graham, F. L., Smiley, J., Russell, W. C. & Nairn, R. ( 1977; ). Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36, 59–74.[CrossRef]
    [Google Scholar]
  10. Griffiths, A. D. & Duncan, A. R. ( 1998; ). Strategies for selection of antibodies by phage display. Curr Opin Biotechnol 9, 102–108.[CrossRef]
    [Google Scholar]
  11. Hanahan, D. & Gluzman, Y. ( 1984; ). Rescue of functional replication origins from embedded configurations in a plasmid carrying the adenovirus genome. Mol Cell Biol 4, 302–309.
    [Google Scholar]
  12. Hatanaka, K., Ohnami, S., Yoshida, K. & 7 other authors ( 2003; ). A simple and efficient method for constructing an adenoviral cDNA expression library. Mol Ther 8, 158–166.[CrossRef]
    [Google Scholar]
  13. Hay, J. G., Shapiro, N., Sauthoff, H., Heitner, S., Phupakdi, W. & Rom, W. N. ( 1999; ). Targeting the replication of adenoviral gene therapy vectors to lung cancer cells: the importance of the adenoviral E1b-55kD gene. Hum Gene Ther 10, 579–590.[CrossRef]
    [Google Scholar]
  14. Hollstein, M., Sidransky, D., Vogelstein, B. & Harris, C. C. ( 1991; ). p53 mutations in human cancers. Science 253, 49–53.[CrossRef]
    [Google Scholar]
  15. Jones, N. & Shenk, T. ( 1979; ). An adenovirus type 5 early gene function regulates expression of other early viral genes. Proc Natl Acad Sci U S A 76, 3665–3669.[CrossRef]
    [Google Scholar]
  16. Ketner, G., Spencer, F., Tugendreich, S., Connelly, C. & Hieter, P. ( 1994; ). Efficient manipulation of the human adenovirus genome as an infectious yeast artificial chromosome clone. Proc Natl Acad Sci U S A 91, 6186–6190.[CrossRef]
    [Google Scholar]
  17. Kitamura, T. ( 1998; ). New experimental approaches in retrovirus-mediated expression screening. Int J Hematol 67, 351–359.[CrossRef]
    [Google Scholar]
  18. Kitchingman, G. R., Lai, S. P. & Westphal, H. ( 1977; ). Loop structures in hybrids of early RNA and the separated strands of adenovirus DNA. Proc Natl Acad Sci U S A 74, 4392–4395.[CrossRef]
    [Google Scholar]
  19. Lochmuller, H., Jani, A., Huard, J., Prescott, S., Simoneau, M., Massie, B., Karpati, G. & Acsadi, G. ( 1994; ). Emergence of early region 1-containing replication-competent adenovirus in stocks of replication-defective adenovirus recombinants (ΔE1+ΔE3) during multiple passages in 293 cells. Hum Gene Ther 5, 1485–1491.[CrossRef]
    [Google Scholar]
  20. Mahlmann, S., McLaughlin, J., Afar, D. E., Mohr, R., Kay, R. J. & Witte, O. N. ( 1998; ). Dissection of signaling pathways and cloning of new signal transducers in tyrosine kinase-induced pathways by genetic selection. Leukemia 12, 1858–1865.[CrossRef]
    [Google Scholar]
  21. McVey, D., Zuber, M., Ettyreddy, D., Brough, D. E. & Kovesdi, I. ( 2002; ). Rapid construction of adenoviral vectors by lambda phage genetics. J Virol 76, 3670–3677.[CrossRef]
    [Google Scholar]
  22. Michiels, F., van Es, H., van Rompaey, L. & 26 other authors ( 2002; ). Arrayed adenoviral expression libraries for functional screening. Nat Biotechnol 20, 1154–1157; erratum 21, 199.
    [Google Scholar]
  23. Miyake, S., Makimura, M., Kanegae, Y., Harada, S., Sato, Y., Takamori, K., Tokuda, C. & Saito, I. ( 1996; ). Efficient generation of recombinant adenoviruses using adenovirus DNA–terminal protein complex and a cosmid bearing the full-length virus genome. Proc Natl Acad Sci U S A 93, 1320–1324.[CrossRef]
    [Google Scholar]
  24. Montell, C., Fisher, E. F., Caruthers, M. H. & Berk, A. J. ( 1982; ). Resolving the functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice site. Nature 295, 380–384.[CrossRef]
    [Google Scholar]
  25. Nevins, J. R. ( 1981; ). Mechanism of activation of early viral transcription by the adenovirus E1A gene product. Cell 26, 213–220.[CrossRef]
    [Google Scholar]
  26. Nevins, J. R., Ginsberg, H. S., Blanchard, J. M., Wilson, M. C. & Darnell, J. E., Jr ( 1979; ). Regulation of the primary expression of the early adenovirus transcription units. J Virol 32, 727–733.
    [Google Scholar]
  27. Shapiro, G. I., Edwards, C. D., Kobzik, L., Godleski, J., Richards, W., Sugarbaker, D. J. & Rollins, B. J. ( 1995; ). Reciprocal Rb inactivation and p16INK4 expression in primary lung cancers and cell lines. Cancer Res 55, 505–509.
    [Google Scholar]
  28. Silhavy, T. J., Berman, M. L. & Enquist, L. W. ( 1984; ). Experiments with Gene Fusions, pp. 303. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  29. Toby, G. G. & Golemis, E. A. ( 2001; ). Using the yeast interaction trap and other two-hybrid-based approaches to study protein–protein interactions. Methods 24, 201–217.[CrossRef]
    [Google Scholar]
  30. Walter, G., Konthur, Z. & Lehrach, H. ( 2001; ). High-throughput screening of surface displayed gene products. Comb Chem High Throughput Screen 4, 193–205.
    [Google Scholar]
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