Abstract
Transcriptional control regions which direct transgene expression to all tissues in transgenic animals can be useful tools for gain-of-function experiments in transgenic animals. A candidate for this purpose is the regulatory region of the human cytomegalovirus immediate early 1 gene (HCMVIE1) which is highly expressed in many lines of tissue culture cells. Here we analyzed the activity of the HCMVIE1 enhancer/promoter using a sensitive reporter gene, the bacterial chloramphenicol acetyltransferase (CAT) gene. Three lines of transgenic mice with an intact transgene were established. All 3 lines expressed the transgene in all 28 tissues analyzed; however, levels of expression between the three lines varied up to 100,000 fold. In addition, expression levels in the high expressing line varied over a 10,000 fold continuum, while expression levels between tissues was almost uniform in the lowest expressing line. The transgene was well expressed in the high expressing line; CAT activity in the highest expressing tissues was equivalent to levels previously reported for tissue specific CAT transgenes active only in a limited number of tissues. These data support the utilization of the HCMVIE1 enhancer/promoter as a means of expressing a transgene in all tissues, but indicate that lines with substantially different overall levels of expression may be generated, and that markedly different levels of tissue specific expression may be found when the overall level of transgene expression is high.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Boshart M., Weber F., Jahn G., Dorsch-Häsler K., Fleckenstein B., Schaffner W. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell. 1985 Jun;41(2):521–530. doi: 10.1016/s0092-8674(85)80025-8. [DOI] [PubMed] [Google Scholar]
- Chang Y. N., Crawford S., Stall J., Rawlins D. R., Jeang K. T., Hayward G. S. The palindromic series I repeats in the simian cytomegalovirus major immediate-early promoter behave as both strong basal enhancers and cyclic AMP response elements. J Virol. 1990 Jan;64(1):264–277. doi: 10.1128/jvi.64.1.264-277.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fang X. J., Keating A., de Villiers J., Sherman M. Tissue-specific activity of heterologous viral promoters in primary rat hepatocytes and Hep G2 cells. Hepatology. 1989 Nov;10(5):781–787. doi: 10.1002/hep.1840100506. [DOI] [PubMed] [Google Scholar]
- Fickenscher H., Stamminger T., Rüger R., Fleckenstein B. The role of a repetitive palindromic sequence element in the human cytomegalovirus major immediate early enhancer. J Gen Virol. 1989 Jan;70(Pt 1):107–123. doi: 10.1099/0022-1317-70-1-107. [DOI] [PubMed] [Google Scholar]
- Foecking M. K., Hofstetter H. Powerful and versatile enhancer-promoter unit for mammalian expression vectors. Gene. 1986;45(1):101–105. doi: 10.1016/0378-1119(86)90137-x. [DOI] [PubMed] [Google Scholar]
- Ghazal P., Lubon H., Fleckenstein B., Hennighausen L. Binding of transcription factors and creation of a large nucleoprotein complex on the human cytomegalovirus enhancer. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3658–3662. doi: 10.1073/pnas.84.11.3658. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghazal P., Lubon H., Hennighausen L. Multiple sequence-specific transcription factors modulate cytomegalovirus enhancer activity in vitro. Mol Cell Biol. 1988 Apr;8(4):1809–1811. doi: 10.1128/mcb.8.4.1809. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunninghake G. W., Monick M. M., Liu B., Stinski M. F. The promoter-regulatory region of the major immediate-early gene of human cytomegalovirus responds to T-lymphocyte stimulation and contains functional cyclic AMP-response elements. J Virol. 1989 Jul;63(7):3026–3033. doi: 10.1128/jvi.63.7.3026-3033.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keating A., Horsfall W., Hawley R. G., Toneguzzo F. Effect of different promoters on expression of genes introduced into hematopoietic and marrow stromal cells by electroporation. Exp Hematol. 1990 Feb;18(2):99–102. [PubMed] [Google Scholar]
- Krippl B., Griep A. E., Mahon K. A., Böhnlein E., Gruss P., Westphal H. Expression and amplification in transgenic mice of a polyoma virus mutant regulatory region. Nucleic Acids Res. 1988 Sep 26;16(18):8963–8976. doi: 10.1093/nar/16.18.8963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leonard J., Khillan J. S., Gendelman H. E., Adachi A., Lorenzo S., Westphal H., Martin M. A., Meltzer M. S. The human immunodeficiency virus long terminal repeat is preferentially expressed in Langerhans cells in transgenic mice. AIDS Res Hum Retroviruses. 1989 Aug;5(4):421–430. doi: 10.1089/aid.1989.5.421. [DOI] [PubMed] [Google Scholar]
- Low M. J., Goodman R. H., Ebert K. M. Cryptic human growth hormone gene sequences direct gonadotroph-specific expression in transgenic mice. Mol Endocrinol. 1989 Dec;3(12):2028–2033. doi: 10.1210/mend-3-12-2028. [DOI] [PubMed] [Google Scholar]
- Merlie J. P., Kornhauser J. M. Neural regulation of gene expression by an acetylcholine receptor promoter in muscle of transgenic mice. Neuron. 1989 Apr;2(4):1295–1300. doi: 10.1016/0896-6273(89)90067-6. [DOI] [PubMed] [Google Scholar]
- Muller S. R., Sullivan P. D., Clegg D. O., Feinstein S. C. Efficient transfection and expression of heterologous genes in PC12 cells. DNA Cell Biol. 1990 Apr;9(3):221–229. doi: 10.1089/dna.1990.9.221. [DOI] [PubMed] [Google Scholar]
- Niller H. H., Hennighausen L. Formation of several specific nucleoprotein complexes on the human cytomegalovirus immediate early enhancer. Nucleic Acids Res. 1991 Jul 11;19(13):3715–3721. doi: 10.1093/nar/19.13.3715. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niller H. H., Hennighausen L. Phytohemagglutinin-induced activity of cyclic AMP (cAMP) response elements from cytomegalovirus is reduced by cyclosporine and synergistically enhanced by cAMP. J Virol. 1990 May;64(5):2388–2391. doi: 10.1128/jvi.64.5.2388-2391.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Overbeek P. A., Lai S. P., Van Quill K. R., Westphal H. Tissue-specific expression in transgenic mice of a fused gene containing RSV terminal sequences. Science. 1986 Mar 28;231(4745):1574–1577. doi: 10.1126/science.3006249. [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]
- Reik W., Williams G., Barton S., Norris M., Neuberger M., Surani M. A. Provision of the immunoglobulin heavy chain enhancer downstream of a test gene is sufficient to confer lymphoid-specific expression in transgenic mice. Eur J Immunol. 1987 Apr;17(4):465–469. doi: 10.1002/eji.1830170405. [DOI] [PubMed] [Google Scholar]
- Rosenthal N., Kornhauser J. M., Donoghue M., Rosen K. M., Merlie J. P. Myosin light chain enhancer activates muscle-specific, developmentally regulated gene expression in transgenic mice. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7780–7784. doi: 10.1073/pnas.86.20.7780. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmidt E. V., Christoph G., Zeller R., Leder P. The cytomegalovirus enhancer: a pan-active control element in transgenic mice. Mol Cell Biol. 1990 Aug;10(8):4406–4411. doi: 10.1128/mcb.10.8.4406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stamminger T., Fickenscher H., Fleckenstein B. Cell type-specific induction of the major immediate early enhancer of human cytomegalovirus by cyclic AMP. J Gen Virol. 1990 Jan;71(Pt 1):105–113. doi: 10.1099/0022-1317-71-1-105. [DOI] [PubMed] [Google Scholar]
- Stinski M. F., Roehr T. J. Activation of the major immediate early gene of human cytomegalovirus by cis-acting elements in the promoter-regulatory sequence and by virus-specific trans-acting components. J Virol. 1985 Aug;55(2):431–441. doi: 10.1128/jvi.55.2.431-441.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]