This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kalos, M.
Right arrow Articles by Fournier, R. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kalos, M.
Right arrow Articles by Fournier, R. E.

 Previous Article  |  Next Article 

Mol. Cell. Biol., Jan 1995, 198-207, Vol 15, No. 1
Copyright © 1995, American Society for Microbiology

Position-independent transgene expression mediated by boundary elements from the apolipoprotein B chromatin domain

M Kalos and RE Fournier
Program in Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.

The human apolipoprotein B (apoB) gene resides within a 47.5-kb chromatin domain that is flanked by sequences that bind to the nuclear matrix. These matrix attachment regions (MARs) are boundaries between nuclease-sensitive and -resistant chromatin. As domain boundaries are thought to function as insulator elements, shielding sequences between them from effects of neighboring chromatin, this raised the possibility that the apoB MARs have functions that could be assayed by transfection. To test this possibility, we examined effects of the apoB MARs on transgene expression in transiently and stably transfected rat and human hepatoma cells. The apoB MARs had no effects on expression of transiently transfected reporters, but they altered expression of stably integrated transgenes in dramatic and reproducible ways. Single integrated copies of transgenes that contained the apoB promoter and second intron enhancer, which are sufficient for high-level expression in transient assays, were expressed at low and variable levels in stable transfectant clones. In contrast, transgenes containing the apoB 5' and 3' MARs were expressed at levels nearly 200-fold higher than levels of the minimal reporters in stable transfectants, and expression was position independent. Transgenes that contained the apoB MARs and an additional 3.3 kb of apoB 5' flanking sequence were also expressed in an elevated, position-independent manner. Surprisingly, tandem transgene arrays in multicopy transfectants were transcriptionally inactive. These observations suggest that the apoB MARs function as insulator elements, shielding transgene expression from effects of neighboring chromatin domains.


This article has been cited by other articles:

  • Petrov, A., Allinne, J., Pirozhkova, I., Laoudj, D., Lipinski, M., Vassetzky, Y. S. (2008). A nuclear matrix attachment site in the 4q35 locus has an enhancer-blocking activity in vivo: Implications for the facio-scapulo-humeral dystrophy. Genome Res 18: 39-45 [Abstract] [Full Text]  
  • Namciu, S. J., Fournier, R. E. K. (2004). Human Matrix Attachment Regions Are Necessary for the Establishment but Not the Maintenance of Transgene Insulation in Drosophila melanogaster. Mol. Cell. Biol. 24: 10236-10245 [Abstract] [Full Text]  
  • Magdinier, F., Yusufzai, T. M., Felsenfeld, G. (2004). Both CTCF-dependent and -independent Insulators Are Found between the Mouse T Cell Receptor {alpha} and Dad1 Genes. J. Biol. Chem. 279: 25381-25389 [Abstract] [Full Text]  
  • Gombert, W. M., Farris, S. D., Rubio, E. D., Morey-Rosler, K. M., Schubach, W. H., Krumm, A. (2003). The c-myc Insulator Element and Matrix Attachment Regions Define the c-myc Chromosomal Domain. Mol. Cell. Biol. 23: 9338-9348 [Abstract] [Full Text]  
  • Giraldo, P., Martinez, A., Regales, L., Lavado, A., Garcia-Diaz, A., Alonso, A., Busturia, A., Montoliu, L. (2003). Functional dissection of the mouse tyrosinase locus control region identifies a new putative boundary activity. Nucleic Acids Res 31: 6290-6305 [Abstract] [Full Text]  
  • Ogata, T., Kozuka, T., Kanda, T. (2003). Identification of an Insulator in AAVS1, a Preferred Region for Integration of Adeno-Associated Virus DNA. J. Virol. 77: 9000-9007 [Abstract] [Full Text]  
  • Li, Q., Peterson, K. R., Fang, X., Stamatoyannopoulos, G. (2002). Locus control regions. Blood 100: 3077-3086 [Abstract] [Full Text]  
  • Liebich, I., Bode, J., Reuter, I., Wingender, E. (2002). Evaluation of sequence motifs found in scaffold/matrix-attached regions (S/MARs). Nucleic Acids Res 30: 3433-3442 [Abstract] [Full Text]  
  • Ehrhardt, A., Kay, M. A. (2002). A new adenoviral helper-dependent vector results in long-term therapeutic levels of human coagulation factor IX at low doses in vivo. Blood 99: 3923-3930 [Abstract] [Full Text]  
  • Shera, K. A., Shera, C. A., McDougall, J. K. (2001). Small Tumor Virus Genomes Are Integrated near Nuclear Matrix Attachment Regions in Transformed Cells. J. Virol. 75: 12339-12346 [Abstract] [Full Text]  
  • Rollini, P., Xu, L., Fournier, R. E. K. (2000). Stable expression and cell-specific chromatin structure of human {alpha}1-antitrypsin cosmid transgenes in rat hepatoma cells. Nucleic Acids Res 28: 3605-3614 [Abstract] [Full Text]  
  • Forrester, W. C., Fernández, L. A., Grosschedl, R. (1999). Nuclear matrix attachment regions antagonize methylation-dependent repression of long-range enhancer-promoter interactions. Genes Dev. 13: 3003-3014 [Abstract] [Full Text]  
  • Rollini, P., Namciu, S. J., Marsden, M. D., Fournier, R. E. (1999). Identification and characterization of nuclear matrix-attachment regions in the human serpin gene cluster at 14q32.1. Nucleic Acids Res 27: 3779-3791 [Abstract] [Full Text]  
  • Zorin, I. D., Gerasimova, T. I., Corces, V. G. (1999). The lawc Gene Is a New Member of the trithorax-Group That Affects the Function of the gypsy Insulator of Drosophila. Genetics 152: 1045-1055 [Abstract] [Full Text]  
  • Hetherington, C. J., Kingsley, P. D., Crocicchio, F., Zhang, P., Rabin, M. S., Palis, J., Zhang, D.-E. (1999). Characterization of Human Endotoxin Lipopolysaccharide Receptor CD14 Expression in Transgenic Mice. J. Immunol. 162: 503-509 [Abstract] [Full Text]  
  • Agarwal, M., Austin, T. W., Morel, F., Chen, J., Bohnlein, E., Plavec, I. (1998). Scaffold Attachment Region-Mediated Enhancement of Retroviral Vector Expression in Primary T Cells. J. Virol. 72: 3720-3728 [Abstract] [Full Text]  
  • Namciu, S. J., Blochlinger, K. B., Fournier, R. E. K. (1998). Human Matrix Attachment Regions Insulate Transgene Expression from Chromosomal Position Effects in Drosophila melanogaster. Mol. Cell. Biol. 18: 2382-2391 [Abstract] [Full Text]  
  • Zhong, X.-P., Krangel, M. S. (1997). An enhancer-blocking element between alpha  and delta  gene segments within the human T cell receptor alpha /delta  locus. Proc. Natl. Acad. Sci. USA 94: 5219-5224 [Abstract] [Full Text]  
  • Palla, F., Melfi, R., Anello, L., Di Bernardo, M., Spinelli, G. (1997). Enhancer blocking activity located near the 3' end of the sea urchin early H2A histone gene. Proc. Natl. Acad. Sci. USA 94: 2272-2277 [Abstract] [Full Text]  
  • Sabourin, J.-C., Kern, A.-S., Grégori, C., Porteu, A., Cywiner, C., Châtelet, F.ço.-P., Kahn, A., Pichard, A.-L. (1996). An Intronic Enhancer Essential for Tissue-specific Expression of the Aldolase B Transgenes. J. Biol. Chem. 271: 3469-3473 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»