Abstract
To assess the feasibility of generating functional transgenes directly via homologous recombination between microinjected DNA fragments, three overlapping genomic DNA fragments, together constituting the human serum albumin (hSA) gene, were coinjected into murine zygotes. The resulting transgenic mice were analyzed for structure and expression of the transgene. All transgenic mice carried recombined hSA DNA fragments and 74% contained a reconstituted hSA gene. HSA expression could be detected in liver and serum in most (72%) of these animals. Only correctly sized hSA transcripts were observed. Transgenic hSA could not be distinguished from the human serum-derived protein by radioimmunoassay or Western blotting. The high frequency and accuracy of homologous recombination in murine zygotes reported here allows the efficient generation of relatively large transgenes.
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- Bollag R. J., Waldman A. S., Liskay R. M. Homologous recombination in mammalian cells. Annu Rev Genet. 1989;23:199–225. doi: 10.1146/annurev.ge.23.120189.001215. [DOI] [PubMed] [Google Scholar]
- Brinster R. L., Allen J. M., Behringer R. R., Gelinas R. E., Palmiter R. D. Introns increase transcriptional efficiency in transgenic mice. Proc Natl Acad Sci U S A. 1988 Feb;85(3):836–840. doi: 10.1073/pnas.85.3.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burke D. T., Carle G. F., Olson M. V. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science. 1987 May 15;236(4803):806–812. doi: 10.1126/science.3033825. [DOI] [PubMed] [Google Scholar]
- Folger K. R., Wong E. A., Wahl G., Capecchi M. R. Patterns of integration of DNA microinjected into cultured mammalian cells: evidence for homologous recombination between injected plasmid DNA molecules. Mol Cell Biol. 1982 Nov;2(11):1372–1387. doi: 10.1128/mcb.2.11.1372. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herbst R. S., Friedman N., Darnell J. E., Jr, Babiss L. E. Positive and negative regulatory elements in the mouse albumin enhancer. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1553–1557. doi: 10.1073/pnas.86.5.1553. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krimpenfort P., Rudenko G., Hochstenbach F., Guessow D., Berns A., Ploegh H. Crosses of two independently derived transgenic mice demonstrate functional complementation of the genes encoding heavy (HLA-B27) and light (beta 2-microglobulin) chains of HLA class I antigens. EMBO J. 1987 Jun;6(6):1673–1676. doi: 10.1002/j.1460-2075.1987.tb02416.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Minghetti P. P., Ruffner D. E., Kuang W. J., Dennison O. E., Hawkins J. W., Beattie W. G., Dugaiczyk A. Molecular structure of the human albumin gene is revealed by nucleotide sequence within q11-22 of chromosome 4. J Biol Chem. 1986 May 25;261(15):6747–6757. [PubMed] [Google Scholar]
- Nuijens J. H., Eerenberg-Belmer A. J., Huijbregts C. C., Schreuder W. O., Felt-Bersma R. J., Abbink J. J., Thijs L. G., Hack C. E. Proteolytic inactivation of plasma C1- inhibitor in sepsis. J Clin Invest. 1989 Aug;84(2):443–450. doi: 10.1172/JCI114185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nuijens J. H., Huijbregts C. C., Eerenberg-Belmer A. J., Meijers J. C., Bouma B. N., Hack C. E. Activation of the contact system of coagulation by a monoclonal antibody directed against a neodeterminant in the heavy chain region of human coagulation factor XII (Hageman factor). J Biol Chem. 1989 Aug 5;264(22):12941–12949. [PubMed] [Google Scholar]
- O'Connor M., Peifer M., Bender W. Construction of large DNA segments in Escherichia coli. Science. 1989 Jun 16;244(4910):1307–1312. doi: 10.1126/science.2660262. [DOI] [PubMed] [Google Scholar]
- Palmiter R. D., Brinster R. L. Germ-line transformation of mice. Annu Rev Genet. 1986;20:465–499. doi: 10.1146/annurev.ge.20.120186.002341. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palmiter R. D., Sandgren E. P., Avarbock M. R., Allen D. D., Brinster R. L. Heterologous introns can enhance expression of transgenes in mice. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):478–482. doi: 10.1073/pnas.88.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pinkert C. A., Ornitz D. M., Brinster R. L., Palmiter R. D. An albumin enhancer located 10 kb upstream functions along with its promoter to direct efficient, liver-specific expression in transgenic mice. Genes Dev. 1987 May;1(3):268–276. doi: 10.1101/gad.1.3.268. [DOI] [PubMed] [Google Scholar]
- Rothschild M. A., Oratz M., Schreiber S. S. Serum albumin. Hepatology. 1988 Mar-Apr;8(2):385–401. doi: 10.1002/hep.1840080234. [DOI] [PubMed] [Google Scholar]
- Sternberg N. Bacteriophage P1 cloning system for the isolation, amplification, and recovery of DNA fragments as large as 100 kilobase pairs. Proc Natl Acad Sci U S A. 1990 Jan;87(1):103–107. doi: 10.1073/pnas.87.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urano Y., Sakai M., Watanabe K., Tamaoki T. Tandem arrangement of the albumin and alpha-fetoprotein genes in the human genome. Gene. 1984 Dec;32(3):255–261. doi: 10.1016/0378-1119(84)90001-5. [DOI] [PubMed] [Google Scholar]
- Urano Y., Watanabe K., Sakai M., Tamaoki T. The human albumin gene. Characterization of the 5' and 3' flanking regions and the polymorphic gene transcripts. J Biol Chem. 1986 Mar 5;261(7):3244–3251. [PubMed] [Google Scholar]
- Wong E. A., Capecchi M. R. Analysis of homologous recombination in cultured mammalian cells in transient expression and stable transformation assays. Somat Cell Mol Genet. 1986 Jan;12(1):63–72. doi: 10.1007/BF01560728. [DOI] [PubMed] [Google Scholar]