Modified GFAP promoter auto-regulates tet-activator expression for increased transactivation and reduced tTA-associated toxicity
Abstract
Transactivator tTA is a necessary component of the tetracycline-regulated inducible gene system. While several transgenic animals have been described that express tTA in the central nervous system (CNS), their tTA levels are often limited, presumably due to toxic effects. We evaluated methods for auto-regulating tTA levels in astrocytes by modifying the transgenic promoter human GFAP (hGFAP). The hGFAP promoter carrying a single copy of the tet-operon in place of a native enhancer element (GFAPtetO1) drove expression of tTA at low levels during un-stimulated, basal condition. However the same promoter auto-induced expression of tTA to significant levels after tetracycline withdrawal. Glial cell-specificity of the promoter remained uncompromised during both basal and induced conditions. Transgenic rats were developed using the auto-inducible GFAPtetO1 promoter that expressed tTA mRNA to high levels in the brain. Expression was widespread within the CNS but enriched in astrocyte-rich regions including the cerebellum. Primary cerebellar astrocytes from GFAPtetO1 rats transfected with 07LacZ produced substantially greater inducibility of reporter gene compared to GFAP-tTA transgenic rats. Finally, GFAPtetO1 rats exhibited severe motor/gait deficit when bred to homozygosity. This phenotype was attributable to developmental abnormalities of the cerebellum and was completely abrogated by doxycycline administration. These results suggest that developmental toxicity resulting from tTA…
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The severity of toxic side-effects of tTA and rtTA depends on the genetic background of the animal and possibly also on the (r)tTA version used.
“The severity of these effects depends on the genetic background of the animal and possibly also on the (r)tTA version used.”
From Tet-On Systems For Doxycycline-inducible Gene Expression (2016) · cites this paper as [80]
In transgenic mice, toxic side-effects of tTA and rtTA have been reported.
“In transgenic mice, toxic side-effects of tTA and rtTA have also been reported [79-83].”
From Tet-On Systems For Doxycycline-inducible Gene Expression (2016) · cites this paper as [80]
Cites
Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.
Gossen, M., & Bujard, H. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.12.5547 · Cited by 4,943 · see citing works
Transcriptional Activation by Tetracyclines in Mammalian Cells
Gossen, M., Freundlieb, S., Bender, G., Müller, G., Hillen, W., & Bujard, H. (1995) · Science · doi:10.1126/science.7792603 · Cited by 2,488 · see citing works
Control of memory formation through regulated expression of a CaMKII transgene
Mayford, M., Bach, M. E., Huang, Y.-Y., Wang, L., Hawkins, R. D., & Kandel, E. R. (1996) · Science · doi:10.1126/science.274.5293.1678 · Cited by 1,550 · see citing works
GAL4-VP16 is an unusually potent transcriptional activator
Sadowski, I., Ma, J., Triezenberg, S., & Ptashne, M. (1988) · Nature · doi:10.1038/335563a0 · Cited by 1,298 · see citing works
Triezenberg, S., Kingsbury, R., & McKnight, S. (1988) · Genes & Development · doi:10.1101/gad.2.6.718 · Cited by 785 · see citing works
Temporal control of gene expression in transgenic mice by a tetracycline-responsive promoter.
Furth, P. A., St‐Onge, L., Boeger, H., Gruss, P., Gossen, M., Kistner, A., Bujard, H., & Hennighausen, L. G. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.20.9302 · Cited by 768 · see citing works
Neurodegeneration in Lurcher mice caused by mutation in δ2 glutamate receptor gene
Zuo, J., De Jager, P. L., Takahashi, K. A., Jiang, W., Linden, D. J., & Heintz, N. (1997) · Nature · doi:10.1038/42009 · Cited by 554 · see citing works
GFAP promoter directs astrocyte-specific expression in transgenic mice
Brenner, M., Kisseberth, W., Su, Y., Besnard, F., & Messing, A. (1994) · Journal of Neuroscience · doi:10.1523/jneurosci.14-03-01030.1994 · Cited by 485 · see citing works
Shockett, P. E., Difilippantonio, M. J., Hellman, N. E., & Schatz, D. G. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.14.6522 · Cited by 378 · see citing works
Baron, U., Gossen, M., & Bujard, H. (1997) · Nucleic Acids Research · doi:10.1093/nar/25.14.2723 · Cited by 370 · see citing works
Live Astrocytes Visualized by Green Fluorescent Protein in Transgenic Mice
Zhuo, L., Sun, B., Zhang, C.-L., Fine, A., Chiu, S. Y., & Messing, A. (1997) · Developmental Biology · doi:10.1006/dbio.1997.8601 · Cited by 364 · see citing works
Transgenic animals with inducible, targeted gene expression in brain.
Chen, J.-S., Kelz, M., Zeng, G., Sakai, N., Steffen, C., Shockett, P. E., Picciotto, M., Duman, R., & Nestler, E. J. (1998) · Molecular Pharmacology · doi:10.1124/mol.54.3.495 · Cited by 194 · see citing works
Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette.
Hofmann, A., Nolan, G. P., & Blau, H. M. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.11.5185 · Cited by 187 · see citing works
Structure and Transcriptional Regulation of the GFAP Gene
Brenner, M. (1994) · Brain Pathology · doi:10.1111/j.1750-3639.1994.tb00840.x · Cited by 150 · see citing works
Besnard, F., Brenner, M., Nakatani, Y., Chao, R., Purohit, H. J., & Freese, E. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)55145-0 · Cited by 146 · see citing works
Cui, W., Allen, N. D., Skynner, M. J., Gusterson, B. A., & Clark, A. J. (2001) · Glia · doi:10.1002/glia.1061 · Cited by 143 · see citing works
Strathdee, C., McLeod, M. R., & Hall, J. R. (1999) · Gene · doi:10.1016/s0378-1119(99)00045-1 · Cited by 138 · see citing works
Conditional Ablation of Cerebellar Astrocytes in Postnatal Transgenic Mice
Delaney, C. L., Brenner, M. J., & Messing, A. (1996) · Journal of Neuroscience · doi:10.1523/jneurosci.16-21-06908.1996 · Cited by 122 · see citing works
Kwak, S., Patel, P. D., Thompson, R. C., Akil, H., & Watson, S. J. (1993) · Endocrinology · doi:10.1210/endo.133.5.8404687 · Cited by 111 · see citing works
Self-citationCharacterization of human cDNA and genomic clones for glial fibrillary acidic protein
Brenner, M., Lampel, K. A., Nakatani, Y., Mill, J. F., Banner, C., Mearow, K. M., Dohadwala, M., Lipsky, R. H., & Freese, E. (1990) · Molecular Brain Research · doi:10.1016/0169-328x(90)90078-r · Cited by 78 · see citing works
Evaluation of an autoregulatory tetracycline regulated system
Gallia, G. L., & Khalili, K. (1998) · Oncogene · doi:10.1038/sj.onc.1201706 · Cited by 45 · see citing works
Studies on Conditional Gene Expression in the Brain
Jerecı̀ć, J., Single, F. N., KRÜTH, U., Krestel, H. E., Kolhekar, R., Storck, T., Kask, K., Higuchi, M., Sprengel, R., & SEEBURG, P. H. (1999) · Annals of the New York Academy of Sciences · doi:10.1111/j.1749-6632.1999.tb11271.x · Cited by 19 · see citing works
Integration of Tetracycline Regulation into a Cell-specific Transcriptional Enhancer
Rose, S. D., & MacDonald, R. J. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.8.4735 · Cited by 13 · see citing works
Doughty, M. L., Lohof, A. M., Selimi, F., Delhaye‐Bouchaud, N., & Mariani, J. (1999) · Journal of Neuroscience · doi:10.1523/jneurosci.19-09-03448.1999 · Cited by 13 · see citing works
Nolte (2001) · doi:10.1002/1098-1136(20010101)33:1
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T. Das, A., Tenenbaum, L., & Berkhout, B. (2016) · Current Gene Therapy · doi:10.2174/1566523216666160524144041 · Cited by 434 · see citing works
“In transgenic mice, toxic side-effects of tTA and rtTA have also been reported [79-83].”
BackgroundPractical considerations for choosing a mouse model of Alzheimer’s disease
Jankowsky, J. L., & Zheng, H. (2017) · Molecular Neurodegeneration · doi:10.1186/s13024-017-0231-7 · Cited by 449 · see citing works
Astrocytes contribute to gamma oscillations and recognition memory
Lee, H. S., Ghetti, A., Pinto‐Duarte, A., Wang, X., Dziewczapolski, G., Galimi, F., Huitrón‐Reséndiz, S., Piña-Crespo, J., Roberts, A. J., Verma, I. M., et al. (2014) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1410893111 · Cited by 259 · see citing works
Neuronal Transgene Expression in Dominant-Negative SNARE Mice
Fujita, T., Chen, M. J., Li, B., Smith, N. A., Peng, W., Sun, W., Toner, M. J., Kress, B. T., Wang, L., Benraiss, A., et al. (2014) · The Journal of Neuroscience · doi:10.1523/jneurosci.2585-14.2014 · Cited by 141 · see citing works
Astrocyte activation and dysfunction and neuron death by HIV-1 Tat expression in astrocytes
Zhou, B. Y., Liu, Y., Kim, B. O., Xiao, Y., & He, J. (2004) · Molecular and Cellular Neuroscience · doi:10.1016/j.mcn.2004.07.003 · Cited by 133 · see citing works
Expression Specificity of GFAP Transgenes
Su, M., Hu, H., Lee, Y., D’Azzo, A., Messing, A., & Brenner, M. (2004) · Neurochemical Research · doi:10.1007/s11064-004-6881-1 · Cited by 124 · see citing works
Han, H. J., Allen, C. C., Buchovecky, C. M., Yetman, M. J., Born, H. A., Marin, M. A., Rodgers, S. P., Song, B. J., Lu, H.-C., Justice, M. J., et al. (2012) · Journal of Neuroscience · doi:10.1523/jneurosci.0893-12.2012 · Cited by 105 · see citing works
Using conditional mutagenesis to study the brain
Morozov, A., Kellendonk, C., Simpson, E. H., & Tronche, F. (2003) · Biological Psychiatry · doi:10.1016/s0006-3223(03)00467-0 · Cited by 72 · see citing works
Tracking progressive pathological and functional decline in the rTg4510 mouse model of tauopathy
Blackmore, T., Meftah, S., Murray, T. K., Craig, P. J., Blockeel, A., Phillips, K., Eastwood, B., O’Neill, M. J., Marston, H., Ahmed, Z., et al. (2017) · Alzheimer's Research & Therapy · doi:10.1186/s13195-017-0306-2 · Cited by 66 · see citing works
Hong, H., Chong, J. L., Song, W., Song, E. J., Jyawook, A. A., Schook, A. C., Ko, C. H., & Takahashi, J. S. (2005) · PLoS Genetics · doi:10.1371/journal.pgen.0030033 · Cited by 59 · see citing works
The power of reversibility regulating gene activities via tetracycline-controlled transcription.
Schönig, K., Bujard, H., & Gossen, M. (2010) · Methods in Enzymology · doi:10.1016/s0076-6879(10)77022-1 · Cited by 45 · see citing works
Matsuzaki, Y., Oue, M., & Hirai, H. (2013) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2013.12.004 · Cited by 32 · see citing works
Reichenstein, M., Rauner, G., & Barash, I. (2011) · Molecular Reproduction and Development · doi:10.1002/mrd.21345 · Cited by 31 · see citing works
Developing tTA Transgenic Rats for Inducible and Reversible Gene Expression
Zhou, H., Huang, C., Yang, M., Landel, C. P., Xia, P. Y., Liu, Y., & Xia, X. (2009) · International Journal of Biological Sciences · doi:10.7150/ijbs.5.171 · Cited by 29 · see citing works
Decreased locomotor activity in mice expressing tTA under control of the CaMKIIα promoter
McKinney, B. C., Schneider, J. S., Schafer, G. L., Lowing, J. L., Mohan, S., Zhao, M., Heng, M. Y., Albin, R. L., Seasholtz, A. F., Akil, H., et al. (2007) · Genes Brain & Behavior · doi:10.1111/j.1601-183x.2007.00339.x · Cited by 25 · see citing works
Conditional mutant mice using tetracycline-controlled gene expression system in the brain
Aiba, A., & Nakao, H. (2007) · Neuroscience Research · doi:10.1016/j.neures.2007.01.009 · Cited by 24 · see citing works
Lent-On-Plus Lentiviral vectors for conditional expression in human stem cells
Benabdellah, K., Muñoz, P., Cobo, M., Gutiérrez-Guerrero, A., Sánchez-Hernández, S., García-Pérez, A., Anderson, P. O., Carrillo-Gálvez, A. B., Toscano, M. G., & Martı́n, F. (2016) · Scientific Reports · doi:10.1038/srep37289 · Cited by 19 · see citing works
Genetic Manipulation in Nutrition, Metabolism, and Obesity Research
Campión, J., Milagro, F. I., & Martínez, J. A. (2004) · Nutrition Reviews · doi:10.1111/j.1753-4887.2004.tb00057.x · Cited by 19 · see citing works
Chew, L., Deboy, C. A., & Senatorov, V. (2014) · Journal of Neuroscience Methods · doi:10.1016/j.jneumeth.2014.08.017 · Cited by 13 · see citing works
Deviation of islet autoreactivity to cryptic epitopes protects NOD mice from diabetes
Song, A., Winer, S., Tsui, H., Sampson, A., Pasceri, P., Ellis, J., Elliott, J. F., & Dosch, H. ‐. (2003) · European Journal of Immunology · doi:10.1002/immu.200310031 · Cited by 12 · see citing works
Tet system in the brain: Transgenic rats and lentiviral vectors approach
Konopka, W., Duniec, K., Klejman, A., Wawrzyniak, M., Owczarek, D., Gawryś, L., Maleszewski, M., Mallet, J. J., & Kaczmarek, L. (2009) · genesis · doi:10.1002/dvg.20487 · Cited by 7 · see citing works
Chyung, Y. H., Peng, P. D., & Kay, M. A. (2003) · Human Gene Therapy · doi:10.1089/104303403767740795 · Cited by 6 · see citing works
Heindorf, M., & Hasan, M. T. (2015) · Cold Spring Harbor Protocols · doi:10.1101/pdb.prot087627 · Cited by 1 · see citing works
- Authors
- Barton, M. D., Dunlop, J. W., Psaltis, G., Kulik, J., DeGennaro, L., & Kwak, S. P.
- Venue
- Molecular Brain Research
- Publisher
- Elsevier BV
- Published
- 2002-05
- Volume
- 101
- Issue
- 1-2
- Pages
- 71-81
- ISSN
- 0169-328X (ISSN-L)
- OpenAlex
- W2094382378
- Cited by
- 23
- References
- 25
- Citations by year
- 2017: 2 · 2016: 2 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 1
- FWCI
- 1.06
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Molecular Brain Research (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- TTA, GFAP promoter, autoregulation, astrocytes, cerebellum, transgenic rats, developmental toxicity, motor impairment, doxycycline, inducible gene expression, tissue-specific gene expression, central nervous system
- MeSH terms
- Animals, Animals, Newborn, Astrocytes, Ataxia, Cell Line, Transformed, Central Nervous System, Enhancer Elements, Genetic, Female, Glial Fibrillary Acidic Protein, Homeostasis, Male, Nervous System Malformations, Phenotype, Promoter Regions, Genetic, Repressor Proteins, RNA, Messenger, Transcriptional Activation, Trans-Activators, Cell Death, Rats, Sprague-Dawley, Gene Expression Regulation, Developmental, Animals, Genetically Modified, Rats
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