A chimeric mammalian transactivator based on the lac repressor that is regulated by temperature and isopropyl beta-D-thiogalactopyranoside.

Baim, S B; Labow, Mark; Levine, Arnold J.; Shenk, Thomas

doi:10.1073/pnas.88.12.5072

Cited by 91 publications

(57 citation statements)

References 38 publications

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“…Several promoter systems are available which are capable of regulating gene expression in eukaryotic cells. These include promoters whose activity is altered in response to heavy metal ions, 13,14 hormones, [15][16][17][18] isopropyl-␤-d-thiogalactoside, 19 tetracycline 5 or to RU486. 6 The metallothionein promoter, the tetracycline (tet) inducible system and a tripartite system based on ecdysone have been tested in transgenic animals 14,18,[20][21][22][23][24] and the tet system has been used in a HSV amplicon vector to regulate reporter gene expression in brain.…”

Section: Discussionmentioning

confidence: 99%

Drug inducible transgene expression in brain using a herpes simplex virus vector

Oligino

Wang

et al. 1998

Gene Ther

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The ability to regulate transgene expression is likely to be ture half maximal expression was achieved with 10important in the use of gene transfer to treat diseases of RU486, and maximal expression was achieved by 24 h. the central nervous system (CNS). In order to achieve Following stereotactic inoculation of the vector into rat regulatable gene expression we created a replicationhippocampus, expression was increased 150-fold by i.p. incompetent genomic herpes simplex vector containing a administration of RU486. This demonstrates that the RU486-inducible transactivator and a lacZ reporter gene RU486 system functions as a tight on/off switch for regunder transcriptional control of a minimal promoter.ulating expression of a transgene delivered to the brain Reporter gene expression from the vector was regulated via an HSV vector. by administration of RU486 in vitro and in vivo. In cell cul-

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Section: Discussionmentioning

confidence: 99%

Drug inducible transgene expression in brain using a herpes simplex virus vector

Oligino

Wang

et al. 1998

Gene Ther

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“…Removing its repressor function and fusing its remaining IPTG-and DNA-binding domains to the VP16 activation domain created a modified version of lacI. This version of lacI functions by an activation mechanism; it binds to lacO sequences when IPTG is present (Deuschle 1989;Baim et al 1991). This chimeric regulator binds lacOcontaining promoters in response to IPTG, thus inducing the expression of a downstream reporter gene.…”

Section: Optimization Of An Inducible System From the Past: The Lactomentioning

confidence: 99%

“…One of the intrinsic difficulties of this system is the problem of generating and maintaining high intracellular concentrations of the repressor protein. Although this system was able to achieve a dramatic induction of reporter gene expression in response to IPTG treatment in mouse cells (Baim et al 1991), inducible gene expression in vivo has been problematic.…”

Section: Optimization Of An Inducible System From the Past: The Lactomentioning

confidence: 99%

Changing colors in mice: an inducible system that delivers: Figure 1.

Mills¹

2001

Genes Dev.

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“…LAP3 cells (Pestov and Lau, 1994), an NIH3T3 cells-derived cell line which constitutively expresses the IPTG-regulated transactivator protein LAP267 (Baim et al, 1991), were maintained in Dulbecco's modi®ed Eagle's medium (DMEM) containing 10% calf serum (Hyclone) and penicillin-streptomycin (GIBCO-BRL). Cells were transfected by the calcium phosphate coprecipitation method (Chen and Okayama, 1988) using 2.5 mg of library DNA or 0.5 mg of a speci®c construct, 0.5 mg of pHyg (Sugden et al, 1985 ) and 2.5 ± 4.5 mg of carrier NIH3T3 DNA per 60-mm dish.…”

Section: Cell Culturementioning

confidence: 99%

Isolation of growth suppressors from a cDNA expression library

1998

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We describe an experimental procedure for the isolation of growth inhibitory sequences from a complex cDNA library. This approach ®rst takes advantage of the SETGAP technique (selectable expression of transient growth arrest phenotype) to enrich for growth inhibitory sequences, followed by a screening procedure to identify individual cDNAs that inhibit cell proliferation. Here we provide a detailed description of the experimental protocol and report the characterization of two cDNA sequences isolated in our initial screen of a mouse cDNA library. One of these cDNAs encodes the mouse ubiquitin-conjugation enzyme UbcM2. The other encodes a truncated form of a novel WD40 repeat protein, named Bop1, which is conserved from yeast to human. Together, these results demonstrate a new approach for the isolation of growth suppressors from cDNA libraries, and identify a previously unknown gene likely to be involved in growth control.

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A chimeric mammalian transactivator based on the lac repressor that is regulated by temperature and isopropyl beta-D-thiogalactopyranoside.

Cited by 91 publications

References 38 publications

Drug inducible transgene expression in brain using a herpes simplex virus vector

Drug inducible transgene expression in brain using a herpes simplex virus vector

Changing colors in mice: an inducible system that delivers: Figure 1.

Isolation of growth suppressors from a cDNA expression library

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