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Proc Natl Acad Sci U S A. Dec 1980; 77(12): 7380–7384.
PMCID: PMC350507

Genetic transformation of mouse embryos by microinjection of purified DNA.

Abstract

A recombinant plasmid composed of segments of herpes simplex virus and simian virus 40 viral DNA inserted into the bacterial plasmid pBR322 was microinjected into pronuclei of fertilized mouse oocytes. The embryos were implanted in the oviducts of pseudopregnant females and allowed to develop to term. DNA from newborn mice was evaluated by the Southern blotting technique for the presence of DNA homologous to the injected plasmid. Two of 78 mice in one series of injections showed clear homology, though the injected sequences had been rearranged. Band intensities from the two positive mice were consistent with the presence of donor DNA in most or all of the cells of the newborns. These results demonstrate that genes can be introduced into the mouse genome by direct insertion into the nuclei of early embryos. This technique affords the opportunity to study problems of gene regulation and cell differentiation in a mammalian system by application of recombinant DNA technology.

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Selected References

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  • Brinster RL. The effect of cells transferred into the mouse blastocyst on subsequent development. J Exp Med. 1974 Oct 1;140(4):1049–1056. [PMC free article] [PubMed]
  • Mintz B, Illmensee K. Normal genetically mosaic mice produced from malignant teratocarcinoma cells. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3585–3589. [PMC free article] [PubMed]
  • Papaioannou VE, McBurney MW, Gardner RL, Evans MJ. Fate of teratocarcinoma cells injected into early mouse embryos. Nature. 1975 Nov 6;258(5530):70–73. [PubMed]
  • Illmensee K, Hoppe PC, Croce CM. Chimeric mice derived from human-mouse hybrid cells. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1914–1918. [PMC free article] [PubMed]
  • Pellicer A, Wagner EF, el-Kareh A, Dewey MJ, Reuser AJ, Silverstein S, Axel R, Mintz B. Introduction of a viral thymidine kinase gene and the human beta-globin gene into developmentally multipotential mouse teratocarcinoma cells. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2098–2102. [PMC free article] [PubMed]
  • Jaenisch R, Mintz B. Simian virus 40 DNA sequences in DNA of healthy adult mice derived from preimplantation blastocysts injected with viral DNA. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1250–1254. [PMC free article] [PubMed]
  • Mullen RJ, Whitten WK. Relationship of genotype and degree of chimerism in coat color to sex ratios and gametogenesis in chimeric mice. J Exp Zool. 1971 Oct;178(2):165–176. [PubMed]
  • Blin N, Stafford DW. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. [PMC free article] [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]
  • Wahl GM, Stern M, Stark GR. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. [PMC free article] [PubMed]
  • Reddy VB, Thimmappaya B, Dhar R, Subramanian KN, Zain BS, Pan J, Ghosh PK, Celma ML, Weissman SM. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. [PubMed]
  • Fiers W, Contreras R, Haegemann G, Rogiers R, Van de Voorde A, Van Heuverswyn H, Van Herreweghe J, Volckaert G, Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. [PubMed]
  • Enquist LW, Vande Woude GF, Wagner M, Smiley JR, Summers WC. Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of Herpes simplex type 1 virus. Gene. 1979 Nov;7(3-4):335–342. [PubMed]
  • Ullrich A, Shine J, Chirgwin J, Pictet R, Tischer E, Rutter WJ, Goodman HM. Rat insulin genes: construction of plasmids containing the coding sequences. Science. 1977 Jun 17;196(4296):1313–1319. [PubMed]
  • Gergen JP, Stern RH, Wensink PC. Filter replicas and permanent collections of recombinant DNA plasmids. Nucleic Acids Res. 1979 Dec 20;7(8):2115–2136. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • D'Eustachio P, Pravtcheva D, Marcu K, Ruddle FH. Chromosomal location of the structural gene cluster encoding murine immunoglobulin heavy chains. J Exp Med. 1980 Jun 1;151(6):1545–1550. [PMC free article] [PubMed]
  • Wigler M, Pellicer A, Silverstein S, Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. [PubMed]
  • Lester SC, LeVan SK, Steglich C, DeMars R. Expression of human genes for adenine phosphoribosyltransferase and hypoxanthine-guanine phosphoribosyltransferase after genetic transformation of mouse cells with purified human DNA. Somatic Cell Genet. 1980 Mar;6(2):241–259. [PubMed]
  • Anderson WF, Killos L, Sanders-Haigh L, Kretschmer PJ, Diacumakos EG. Replication and expression of thymidine kinase and human globin genes microinjected into mouse fibroblasts. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5399–5403. [PMC free article] [PubMed]
  • ter Schegget J, Voves J, van Strien A, van der Noordaa J. Free viral DNA in BK virus-induced hamster tumor cells. J Virol. 1980 Aug;35(2):331–339. [PMC free article] [PubMed]
  • Hanahan D, Lane D, Lipsich L, Wigler M, Botchan M. Characteristics of an SV40-plasmid recombinant and its movement into and out of the genome of a murine cell. Cell. 1980 Aug;21(1):127–139. [PubMed]
  • Kelly TJ, Jr, Lewis AM, Jr, Levine AS, Siegel S. Structure of two adenovirus-simian virus 40 hybrids which contain the entire SV40 genome. J Mol Biol. 1974 Oct 15;89(1):113–126. [PubMed]
  • Botchan M, Topp W, Sambrook J. Studies on simian virus 40 excision from cellular chromosomes. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):709–719. [PubMed]
  • Huttner KM, Scangos GA, Ruddle FH. DNA-mediated gene transfer of a circular plasmid into murine cells. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5820–5824. [PMC free article] [PubMed]

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