• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Feb 1988; 85(3): 836–840.
PMCID: PMC279650

Introns increase transcriptional efficiency in transgenic mice.

Abstract

Experiments were designed to test the effect of introns on gene expression in transgenic mice. Four different pairs of gene constructs, which were identical except that one member of each pair lacked all introns, were compared for expression of mRNA after introduction into the murine germ line by microinjection of fertilized eggs. The expression of two chimeric genes, made by fusing either the mouse metallothionein I or the rat elastase 1 promoter/enhancer to the rat growth hormone gene, was assayed in fetal liver or pancreas, respectively, while two natural genes, an oligonucleotide-marked mouse metallothionein I gene and the human beta-globin gene, were assayed in fetal liver. In each case there was, on average, 10- to 100-fold more mRNA produced from the intron-containing construct. Moreover, mRNA levels were proportional to the relative rates of transcription that were measured in isolated nuclei. However, when the expression of the two mouse metallothionein I gene-based constructs was tested after transfection into cultured cells, little difference was observed. These observations suggest that introns play a role in facilitating transcription of microinjected genes and that this effect may be manifest only on genes exposed to developmental influences.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Gilbert W. Why genes in pieces? Nature. 1978 Feb 9;271(5645):501–501. [PubMed]
  • Südhof TC, Russell DW, Goldstein JL, Brown MS, Sanchez-Pescador R, Bell GI. Cassette of eight exons shared by genes for LDL receptor and EGF precursor. Science. 1985 May 17;228(4701):893–895. [PubMed]
  • Breitbart RE, Nguyen HT, Medford RM, Destree AT, Mahdavi V, Nadal-Ginard B. Intricate combinatorial patterns of exon splicing generate multiple regulated troponin T isoforms from a single gene. Cell. 1985 May;41(1):67–82. [PubMed]
  • Leff SE, Rosenfeld MG, Evans RM. Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem. 1986;55:1091–1117. [PubMed]
  • Hamer DH, Leder P. Splicing and the formation of stable RNA. Cell. 1979 Dec;18(4):1299–1302. [PubMed]
  • Gruss P, Lai CJ, Dhar R, Khoury G. Splicing as a requirement for biogenesis of functional 16S mRNA of simian virus 40. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4317–4321. [PMC free article] [PubMed]
  • Gruss P, Khoury G. Rescue of a splicing defective mutant by insertion of an heterologous intron. Nature. 1980 Aug 7;286(5773):634–637. [PubMed]
  • Mulligan RC, Berg P. Expression of a bacterial gene in mammalian cells. Science. 1980 Sep 19;209(4463):1422–1427. [PubMed]
  • Carlock L, Jones NC. Synthesis of an unspliced cytoplasmic message by an adenovirus 5 deletion mutant. Nature. 1981 Dec 10;294(5841):572–574. [PubMed]
  • Treisman R, Novak U, Favaloro J, Kamen R. Transformation of rat cells by an altered polyoma virus genome expressing only the middle-T protein. Nature. 1981 Aug 13;292(5824):595–600. [PubMed]
  • Chee PP, Klassy RC, Slightom JL. Expression of a bean storage protein 'phaseolin minigene' in foreign plant tissues. Gene. 1986;41(1):47–57. [PubMed]
  • Ng R, Domdey H, Larson G, Rossi JJ, Abelson J. A test for intron function in the yeast actin gene. Nature. 1985 Mar 14;314(6007):183–184. [PubMed]
  • Gross MK, Kainz MS, Merrill GF. Introns are inconsequential to efficient formation of cellular thymidine kinase mRNA in mouse L cells. Mol Cell Biol. 1987 Dec;7(12):4576–4581. [PMC free article] [PubMed]
  • Brinster RL, Palmiter RD. Introduction of genes into the germ line of animals. Harvey Lect. 1984 1985;80:1–38. [PubMed]
  • Palmiter RD, Brinster RL. Germ-line transformation of mice. Annu Rev Genet. 1986;20:465–499. [PubMed]
  • Durnam DM, Perrin F, Gannon F, Palmiter RD. Isolation and characterization of the mouse metallothionein-I gene. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6511–6515. [PMC free article] [PubMed]
  • Barta A, Richards RI, Baxter JD, Shine J. Primary structure and evolution of rat growth hormone gene. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4867–4871. [PMC free article] [PubMed]
  • Ornitz DM, Palmiter RD, Hammer RE, Brinster RL, Swift GH, MacDonald RJ. Specific expression of an elastase-human growth hormone fusion gene in pancreatic acinar cells of transgenic mice. Nature. 1985 Feb 14;313(6003):600–602. [PubMed]
  • Glanville N, Durnam DM, Palmiter RD. Structure of mouse metallothionein-I gene and its mRNA. Nature. 1981 Jul 16;292(5820):267–269. [PubMed]
  • Collins FS, Weissman SM. The molecular genetics of human hemoglobin. Prog Nucleic Acid Res Mol Biol. 1984;31:315–462. [PubMed]
  • Brinster RL, Chen HY, Trumbauer ME, Yagle MK, Palmiter RD. Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4438–4442. [PMC free article] [PubMed]
  • Palmiter RD, Chen HY, Brinster RL. Differential regulation of metallothionein-thymidine kinase fusion genes in transgenic mice and their offspring. Cell. 1982 Jun;29(2):701–710. [PubMed]
  • Townes TM, Lingrel JB, Chen HY, Brinster RL, Palmiter RD. Erythroid-specific expression of human beta-globin genes in transgenic mice. EMBO J. 1985 Jul;4(7):1715–1723. [PMC free article] [PubMed]
  • Durnam DM, Palmiter RD. A practical approach for quantitating specific mRNAs by solution hybridization. Anal Biochem. 1983 Jun;131(2):385–393. [PubMed]
  • Mulvihill ER, Palmiter RD. Relationship of nuclear estrogen receptor levels to induction of ovalbumin and conalbumin mRNA in chick oviduct. J Biol Chem. 1977 Mar 25;252(6):2060–2068. [PubMed]
  • McKnight GS, Palmiter RD. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed]
  • Searle PF, Stuart GW, Palmiter RD. Building a metal-responsive promoter with synthetic regulatory elements. Mol Cell Biol. 1985 Jun;5(6):1480–1489. [PMC free article] [PubMed]
  • Hamer DH. Metallothionein. Annu Rev Biochem. 1986;55:913–951. [PubMed]
  • Palmiter RD, Norstedt G, Gelinas RE, Hammer RE, Brinster RL. Metallothionein-human GH fusion genes stimulate growth of mice. Science. 1983 Nov 18;222(4625):809–814. [PubMed]
  • Hammer RE, Swift GH, Ornitz DM, Quaife CJ, Palmiter RD, Brinster RL, MacDonald RJ. The rat elastase I regulatory element is an enhancer that directs correct cell specificity and developmental onset of expression in transgenic mice. Mol Cell Biol. 1987 Aug;7(8):2956–2967. [PMC free article] [PubMed]
  • Banerji J, Olson L, Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. [PubMed]
  • Gillies SD, Morrison SL, Oi VT, Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. [PubMed]
  • Behringer RR, Hammer RE, Brinster RL, Palmiter RD, Townes TM. Two 3' sequences direct adult erythroid-specific expression of human beta-globin genes in transgenic mice. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7056–7060. [PMC free article] [PubMed]
  • Stuart GW, Searle PF, Chen HY, Brinster RL, Palmiter RD. A 12-base-pair DNA motif that is repeated several times in metallothionein gene promoters confers metal regulation to a heterologous gene. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7318–7322. [PMC free article] [PubMed]
  • Nelson C, Crenshaw EB, 3rd, Franco R, Lira SA, Albert VR, Evans RM, Rosenfeld MG. Discrete cis-active genomic sequences dictate the pituitary cell type-specific expression of rat prolactin and growth hormone genes. Nature. 1986 Aug 7;322(6079):557–562. [PubMed]
  • Lefevre C, Imagawa M, Dana S, Grindlay J, Bodner M, Karin M. Tissue-specific expression of the human growth hormone gene is conferred in part by the binding of a specific trans-acting factor. EMBO J. 1987 Apr;6(4):971–981. [PMC free article] [PubMed]
  • Eissenberg JC, Cartwright IL, Thomas GH, Elgin SC. Selected topics in chromatin structure. Annu Rev Genet. 1985;19:485–536. [PubMed]
  • Richard-Foy H, Hager GL. Sequence-specific positioning of nucleosomes over the steroid-inducible MMTV promoter. EMBO J. 1987 Aug;6(8):2321–2328. [PMC free article] [PubMed]
  • Chambon P. Summary: the molecular biology of the eukaryotic genome is coming of age. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):1209–1234. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...