Home LiteratureArticle Details
PMID: 10448644 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro.

Molecular medicine (Cambridge, Mass.) ·Vol. 5 ·No. 4 ·1999-04-00 ·Pages 224-31

Aoki K, Barker C, Danthinne X, Imperiale MJ, Nabel GJ

Abstract

Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system. Full-length recombinant adenovirus DNA was generated in vitro by Cre-mediated recombination between loxP sites in a linearized shuttle plasmid containing a transgene and adenovirus genomic DNA. After transfection of Cre-treated DNA into 293 cells, replication-defective viral vectors were rapidly obtained without detectable wild-type virus. This system facilitates the development of recombinant adenoviral vectors for basic and clinical research.

MeSH Terms
Adenoviridae/genetics,growth & development Animals Cells, Cultured Cosmids Cytopathogenic Effect, Viral DNA, Viral/genetics Genes, Reporter Genetic Vectors Integrases/metabolism Lac Operon Muscle, Smooth/cytology Plasmids/genetics Recombination, Genetic Swine Transfection Viral Proteins
Chemicals
DNA, Viral Viral Proteins Cre recombinase Integrases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Aoki K
Howard Hughes Medical Institute, University of Michigan Medical Center, Department of Internal Medicine and Biological Chemistry, Ann Arbor 48109-0650, USA.
Barker C
Danthinne X
Imperiale M J
Nabel G J
References (26)
26 references, click to expand
  1. The infectivity of adenovirus 5 DNA-protein complex.
    Virology. 1976 Dec;75(2):442-56 PMID: 1032904
  2. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting.
    Science. 1994 Jul 1;265(5168):103-6 PMID: 8016642
  3. P1 site-specific recombination: nucleotide sequence of the recombining sites.
    Proc Natl Acad Sci U S A. 1982 Jun;79(11):3398-402 PMID: 6954485
  4. Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination.
    Cell. 1983 Apr;32(4):1301-11 PMID: 6220808
  5. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  6. Interaction of the bacteriophage P1 recombinase Cre with the recombining site loxP.
    Proc Natl Acad Sci U S A. 1984 Feb;81(4):1026-9 PMID: 6230671
  7. Mechanism of strand cleavage and exchange in the Cre-lox site-specific recombination system.
    J Mol Biol. 1985 Feb 5;181(3):351-62 PMID: 3856690
  8. In vitro and in vivo synthesis of the hepatitis B virus surface antigen and of the receptor for polymerized human serum albumin from recombinant human adenoviruses.
    EMBO J. 1985 Dec 30;4(13B):3861-5 PMID: 3004975
  9. A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5.
    Virology. 1988 Apr;163(2):614-7 PMID: 2965450
  10. Expression of human alpha 1-antitrypsin using a recombinant adenovirus vector.
    FEBS Lett. 1990 Jul 2;267(1):60-2 PMID: 2365091
  11. Adenovirus-mediated transfer of a recombinant alpha 1-antitrypsin gene to the lung epithelium in vivo.
    Science. 1991 Apr 19;252(5004):431-4 PMID: 2017680
  12. Widespread long-term gene transfer to mouse skeletal muscles and heart.
    J Clin Invest. 1992 Aug;90(2):626-30 PMID: 1644927
  13. Cre-lox recombination in Escherichia coli cells. Mechanistic differences from the in vitro reaction.
    J Mol Biol. 1992 Aug 5;226(3):661-73 PMID: 1324323
  14. Genomic targeting with a positive-selection lox integration vector allows highly reproducible gene expression in mammalian cells.
    Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7905-9 PMID: 1518811
  15. A model system for in vivo gene transfer into the central nervous system using an adenoviral vector.
    Nat Genet. 1993 Mar;3(3):219-23 PMID: 8387378
  16. Genomic targeting with purified Cre recombinase.
    Nucleic Acids Res. 1993 May 11;21(9):2025-9 PMID: 8502542
  17. Flipping fruit-flies: a powerful new technique for generating Drosophila mosaics.
    Trends Genet. 1993 Jul;9(7):227-8 PMID: 8378996
  18. Site-specific recombinases: tools for genome engineering.
    Trends Genet. 1993 Dec;9(12):413-21 PMID: 8122308
  19. Efficient manipulation of the human adenovirus genome as an infectious yeast artificial chromosome clone.
    Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6186-90 PMID: 8016135
  20. Site-specific recombination mediated by an adenovirus vector expressing the Cre recombinase protein: a molecular switch for control of gene expression.
    J Virol. 1995 Aug;69(8):4600-6 PMID: 7609024
  21. Replicating vectors for gene therapy of cancer: risks, limitations and prospects.
    Eur J Cancer. 1994;30A(8):1165-71 PMID: 7654450
  22. Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli.
    J Virol. 1996 Jul;70(7):4805-10 PMID: 8676512
  23. Recombinational construction in Escherichia coli of infectious adenoviral genomes.
    Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1414-9 PMID: 9037067
  24. Use of the cosmid adenoviral vector cloning system for the in vitro construction of recombinant adenoviral vectors.
    Hum Gene Ther. 1997 Jul 20;8(11):1321-30 PMID: 9295127
  25. A simplified system for generating recombinant adenoviruses.
    Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2509-14 PMID: 9482916
  26. Physical mapping of a large-plaque mutation of adenovirus type 2.
    J Virol. 1979 Nov;32(2):623-8 PMID: 501803
Article Info
Journal
Molecular medicine (Cambridge, Mass.)
Abbr.
Mol Med
ISSN
1076-1551
Published
1999-04-00
Pages
224-31
Language
English
Region
England
NLM ID
9501023
PMCID
PMC2230325
Subset
IM
Grants
NIGMS NIH HHS · GM34902 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2