A tetracycline controlled activation/repression system with increased potential for gene transfer into mammalian cells

Tight control of gene activity has been achieved in cells and transgenic organisms using the Tet regulatory systems. Unregulated basal transcription can, however, be observed whenever integration of target genes driven by promoters responsive to tetracycline controlled transcriptional activators (tTA, rtTA) does not occur at suitable chromosomal sites. Moreover, in viral vectors containing both the tTA coding sequence and the regulated target gene, proximity of the enhancer element driving tTA/rtTA expression to the responsive unit will lead to elevated background levels. Similarly when tTA/rtTA responsive transcription units are in a non‐integrated state as eg., during transient expression, intrinsic residual transcription persists in their ‘off’ state, which can differ in intensity among different cell types.

[1]  Erik T. Bieschke,et al.  Doxycycline-induced transgene expression during Drosophila development and aging , 1998, Molecular and General Genetics MGG.

[2]  M. Gossen,et al.  Control of gene activity in higher eukaryotic cells by prokaryotic regulatory elements. , 1994, Trends in biochemical sciences.

[3]  J. Ortín,et al.  Expression in mammalian cells of a gene from Streptomyces alboniger conferring puromycin resistance. , 1986, Nucleic acids research.

[4]  M. L. Kaplan,et al.  Tetracycline-regulated cardiac gene expression in vivo. , 1994, The Journal of clinical investigation.

[5]  M. Gossen,et al.  Transcriptional activation by tetracyclines in mammalian cells. , 1995, Science.

[6]  J. Manley,et al.  Transcriptional repression by the Drosophila even-skipped protein: definition of a minimal repression domain. , 1993, Genes & development.

[7]  M. Gossen,et al.  Use of tetracycline-controlled gene expression systems to study mammalian cell cycle. , 1997, Methods in enzymology.

[8]  W. Gehring,et al.  In vivo analysis of scaffold‐associated regions in Drosophila: a synthetic high‐affinity SAR binding protein suppresses position effect variegation , 1998, The EMBO journal.

[9]  J. Bonventre,et al.  The Krüppel-associated box-A (KRAB-A) domain of zinc finger proteins mediates transcriptional repression. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[10]  D. Bohl,et al.  Long-term control of erythropoietin secretion by doxycycline in mice transplanted with engineered primary myoblasts , 1997, Nature Medicine.

[11]  U. Deuschle,et al.  Tetracycline-reversible silencing of eukaryotic promoters , 1995, Molecular and cellular biology.

[12]  G. Fishman,et al.  Conditional lineage ablation to model human diseases. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Uney,et al.  Switching transgene expression in the brain using an adenoviral tetracycline-regulatable system , 1998, Nature Biotechnology.

[14]  Michael Boshart,et al.  A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus , 1985, Cell.

[15]  M. Gossen,et al.  Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[16]  M. Gossen,et al.  Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[17]  M. Gossen,et al.  A chimeric transactivator allows tetracycline-responsive gene expression in whole plants. , 1994, The Plant journal : for cell and molecular biology.

[18]  M. Gossen,et al.  Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. , 1997, Nucleic acids research.

[19]  R. Schimke,et al.  Tetracycline-controlled gene expression system achieves high-level and quantitative control of gene expression. , 1996, Analytical biochemistry.

[20]  M Aldea,et al.  A Set of Vectors with a Tetracycline‐Regulatable Promoter System for Modulated Gene Expression in Saccharomyces cerevisiae , 1997, Yeast.

[21]  William D. Richardson,et al.  A short amino acid sequence able to specify nuclear location , 1984, Cell.

[22]  E. Kandel,et al.  Control of Memory Formation Through Regulated Expression of a CaMKII Transgene , 1996, Science.

[23]  M. Gossen,et al.  Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Efrat,et al.  Conditional transformation of a pancreatic beta-cell line derived from transgenic mice expressing a tetracycline-regulated oncogene. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  L. Hennighausen,et al.  Time-Sensitive Reversal of Hyperplasia in Transgenic Mice Expressing SV40 T Antigen , 1996, Science.

[26]  R. Renkawitz,et al.  Two silencing sub-domains of v-erbA synergize with each other, but not with RXR. , 1994, Nucleic acids research.