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Proc Natl Acad Sci U S A. Jul 1988; 85(14): 5166–5170.
PMCID: PMC281709

Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1.


The Cre protein encoded by the coliphage P1 is a 38-kDa protein that efficiently promotes both intra- and intermolecular synapsis and recombination of DNA both in Escherichia coli and in vitro. Recombination occurs at a specific site, called lox, and does not require any other protein factors. The Cre protein is shown here also to be able to cause synapsis of DNA and site-specific recombination in a mammalian cell line. A stable mouse cell line was established that expresses the Cre protein under the control of the Cd2+-inducible metallothionein I gene promoter. DNA recombination was monitored with DNA substrates containing two directly repeated lox sites. One such substrate is a circular plasmid with two directly repeated lox sites (lox2) flanking a marker gene and was introduced into cells by Ca3(PO4)2 transformation. As a second substrate we used a pseudorabies virus (a herpesvirus) containing a lox2 insertion designed to provide a sensitive detection system for recombination. In both cases, site-specific recombination in vivo is dependent on the presence of the Cre protein and occurs specifically at the 34-base-pair lox sites. These results demonstrate the controlled site-specific synapsis of DNA and recombination by a prokaryotic protein in mammalian cells and suggest that Cre-mediated site-specific recombination may be a useful tool for understanding and modulating genome rearrangements in eukaryotes.

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  • Nowell PC, Croce CM. Chromosomes, genes, and cancer. Am J Pathol. 1986 Oct;125(1):7–15. [PMC free article] [PubMed]
  • Sternberg N, Hamilton D. Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites. J Mol Biol. 1981 Aug 25;150(4):467–486. [PubMed]
  • Abremski K, Hoess R, Sternberg N. Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination. Cell. 1983 Apr;32(4):1301–1311. [PubMed]
  • Hoess RH, Abremski K. Mechanism of strand cleavage and exchange in the Cre-lox site-specific recombination system. J Mol Biol. 1985 Feb 5;181(3):351–362. [PubMed]
  • Sauer B. Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jun;7(6):2087–2096. [PMC free article] [PubMed]
  • Pavlakis GN, Hamer DH. Regulation of a metallothionein-growth hormone hybrid gene in bovine papilloma virus. Proc Natl Acad Sci U S A. 1983 Jan;80(2):397–401. [PMC free article] [PubMed]
  • Sauer B, Whealy M, Robbins A, Enquist L. Site-specific insertion of DNA into a pseudorabies virus vector. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9108–9112. [PMC free article] [PubMed]
  • Holland TC, Sandri-Goldin RM, Holland LE, Marlin SD, Levine M, Glorioso JC. Physical mapping of the mutation in an antigenic variant of herpes simplex virus type 1 by use of an immunoreactive plaque assay. J Virol. 1983 May;46(2):649–652. [PMC free article] [PubMed]
  • Lowy DR, Rands E, Scolnick EM. Helper-independent transformation by unintegrated Harvey sarcoma virus DNA. J Virol. 1978 May;26(2):291–298. [PMC free article] [PubMed]
  • Lowy DR, Dvoretzky I, Shober R, Law MF, Engel L, Howley PM. In vitro tumorigenic transformation by a defined sub-genomic fragment of bovine papilloma virus DNA. Nature. 1980 Sep 4;287(5777):72–74. [PubMed]
  • Sarver N, Gruss P, Law MF, Khoury G, Howley PM. Bovine papilloma virus deoxyribonucleic acid: a novel eucaryotic cloning vector. Mol Cell Biol. 1981 Jun;1(6):486–496. [PMC free article] [PubMed]
  • Lin FL, Sperle KM, Sternberg NL. Extrachromosomal recombination in mammalian cells as studied with single- and double-stranded DNA substrates. Mol Cell Biol. 1987 Jan;7(1):129–140. [PMC free article] [PubMed]
  • Scott RW, Vogt TF, Croke ME, Tilghman SM. Tissue-specific activation of a cloned alpha-fetoprotein gene during differentiation of a transfected embryonal carcinoma cell line. Nature. 1984 Aug 16;310(5978):562–567. [PubMed]
  • Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Ben-Porat T, Kaplan AS. Synthesis of proteins in cells infected with herpesvirus. V. Viral glycoproteins. Virology. 1970 Jun;41(2):265–273. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Aviv H, Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. [PMC free article] [PubMed]
  • Abremski K, Hoess R. Bacteriophage P1 site-specific recombination. Purification and properties of the Cre recombinase protein. J Biol Chem. 1984 Feb 10;259(3):1509–1514. [PubMed]
  • Karin M, Herschman HR. Induction of metallothionein in HeLa cells by dexamethasone and zinc. Eur J Biochem. 1981 Jan;113(2):267–272. [PubMed]
  • Abremski K, Hoess R. Phage P1 Cre-loxP site-specific recombination. Effects of DNA supercoiling on catenation and knotting of recombinant products. J Mol Biol. 1985 Jul 20;184(2):211–220. [PubMed]
  • Holland TC, Homa FL, Marlin SD, Levine M, Glorioso J. Herpes simplex virus type 1 glycoprotein C-negative mutants exhibit multiple phenotypes, including secretion of truncated glycoproteins. J Virol. 1984 Nov;52(2):566–574. [PMC free article] [PubMed]

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