Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette.

Abstract:

:We describe a single autoregulatory cassette that allows reversible induction of transgene expression in response to tetracycline (tet). This cassette contains all of the necessary components previously described by others on two separate plasmids that are introduced sequentially over a period of months [Gossen, M. & Bujard, H. (1992) Proc. Natl. Acad. Sci. USA 89, 5547-5551]. The cassette is introduced using a retrovirus, allowing transfer into cell types that are difficult to transfect. Thus, populations of thousands of cells, rather than a few clones, can be isolated and characterized within weeks. To avoid potential interference of the strong retroviral long terminal repeat enhancer and promoter elements with the function of the tet-regulated cytomegalovirus minimal promoter, the vector is self-inactivating, eliminating transcription from the long terminal repeat after infection of target cells. Tandem tet operator sequences and the cytomegalovirus minimal promoter drive expression of a bicistronic mRNA, leading to transcription of the gene of interest (lacZ) and the internal ribosome entry site controlled transactivator (Tet repressor-VP16 fusion protein). In the absence of tet, there is a progressive increase in transactivator by means of an autoregulatory loop, whereas in the presence of tet, gene expression is prevented. Northern blot, biochemical, and single cell analyses have all shown that the construct yields low basal levels of gene expression and induction of one to two orders of magnitude. Thus, the current cassette of the retroviral construct (SIN-RetroTet vector) allows rapid delivery of inducible genes and should have broad applications to cultured cells, transgenic animals, and gene therapy.

authors

Hofmann A,Nolan GP,Blau HM

doi

10.1073/pnas.93.11.5185

subject

Has Abstract

pub_date

1996-05-28 00:00:00

pages

5185-90

issue

11

eissn

0027-8424

issn

1091-6490

journal_volume

93

pub_type

评论,杂志文章
  • Regulation of protein synthesis in reticulocyte lysates: phosphorylation of methionyl-tRNAf binding factor by protein kinase activity of translational inhibitor isolated from hemedeficient lysates.

    abstract::A previous study demonstrated that the translational inhibitor from lysates of heme-deficient rabbit reticulocytes is associated with a protein kinase activity. Chromatography of this inhibitor preparation on phosphocellulose yields two distinct protein kinase activities, PC1 and PC2. PC1, which consitutes about 90% o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.9.3112

    authors: Levin D,Ranu RS,Ernst V,London IM

    更新日期:1976-09-01 00:00:00

  • Low dielectric response in enzyme active site.

    abstract::The kinetics of charge transfer depend crucially on the dielectric reorganization of the medium. In enzymatic reactions that involve charge transfer, atomic dielectric response of the active site and of its surroundings determines the efficiency of the protein as a catalyst. We report direct spectroscopic measurements...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.050316997

    authors: Mertz EL,Krishtalik LI

    更新日期:2000-02-29 00:00:00

  • Tumor-infiltrating NY-ESO-1-specific CD8+ T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer.

    abstract::NY-ESO-1 is a "cancer-testis" antigen frequently expressed in epithelial ovarian cancer (EOC) and is among the most immunogenic tumor antigens defined to date. In an effort to understand in vivo tolerance mechanisms, we assessed the phenotype and function of NY-ESO-1-specific CD8(+) T cells derived from peripheral blo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1003345107

    authors: Matsuzaki J,Gnjatic S,Mhawech-Fauceglia P,Beck A,Miller A,Tsuji T,Eppolito C,Qian F,Lele S,Shrikant P,Old LJ,Odunsi K

    更新日期:2010-04-27 00:00:00

  • Highly accurate two-gene classifier for differentiating gastrointestinal stromal tumors and leiomyosarcomas.

    abstract::Gastrointestinal stromal tumor (GIST) has emerged as a clinically distinct type of sarcoma with frequent overexpression and mutation of the c-Kit oncogene and a favorable response to imatinib mesylate [also known as STI571 (Gleevec)] therapy. However, a significant diagnostic challenge remains in the differentiation o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0611373104

    authors: Price ND,Trent J,El-Naggar AK,Cogdell D,Taylor E,Hunt KK,Pollock RE,Hood L,Shmulevich I,Zhang W

    更新日期:2007-02-27 00:00:00

  • Biological impact on mineral dissolution: application of the lichen model to understanding mineral weathering in the rhizosphere.

    abstract::Microorganisms modify rates and mechanisms of chemical and physical weathering and clay growth, thus playing fundamental roles in soil and sediment formation. Because processes in soils are inherently complex and difficult to study, we employ a model based on the lichen-mineral system to identify the fundamental inter...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.7.3404

    authors: Banfield JF,Barker WW,Welch SA,Taunton A

    更新日期:1999-03-30 00:00:00

  • Role of the regulatory subunit of bovine pyruvate dehydrogenase phosphatase.

    abstract::Bovine pyruvate dehydrogenase phosphatase (PDP) is a Mg2+-dependent and Ca2+-stimulated heterodimer that is a member of the protein phosphatase 2C family and is localized to mitochondria. Insight into the function of the regulatory subunit of PDP (PDPr) has been gained. It decreases the sensitivity of the catalytic su...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.10.4953

    authors: Yan J,Lawson JE,Reed LJ

    更新日期:1996-05-14 00:00:00

  • Serial coalescent simulations suggest a weak genealogical relationship between Etruscans and modern Tuscans.

    abstract::The Etruscans, the only preclassical European population that has been genetically characterized so far, share only two haplotypes with their modern geographic counterparts, the Tuscans, who, nonetheless, appear to be their closest relatives. We modeled 10 demographic scenarios spanning the last 2,500 years and tested...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0509718103

    authors: Belle EM,Ramakrishnan U,Mountain JL,Barbujani G

    更新日期:2006-05-23 00:00:00

  • Cultural evolution of conformity and anticonformity.

    abstract::Conformist bias occurs when the probability of adopting a more common cultural variant in a population exceeds its frequency, and anticonformist bias occurs when the reverse is true. Conformist and anticonformist bias have been widely documented in humans, and conformist bias has also been observed in many nonhuman an...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2004102117

    authors: Denton KK,Ram Y,Liberman U,Feldman MW

    更新日期:2020-06-16 00:00:00

  • Carbohydrate self-recognition mediates marine sponge cellular adhesion.

    abstract::Sponges (Porifera), the simplest and earliest multicellular organisms, are thought to have evolved from their unicellular ancestors about 1 billion years ago by developing cell-recognition and adhesion mechanisms to discriminate against "non-self." Consequently, they are used as models for investigating recognition ph...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.151111298

    authors: Haseley SR,Vermeer HJ,Kamerling JP,Vliegenthart JF

    更新日期:2001-07-31 00:00:00

  • Evidence for a recent increase in forest growth.

    abstract::Forests and their soils contain the majority of the earth's terrestrial carbon stocks. Changes in patterns of tree growth can have a huge impact on atmospheric cycles, biogeochemical cycles, climate change, and biodiversity. Recent studies have shown increases in biomass across many forest types. This increase has bee...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0912376107

    authors: McMahon SM,Parker GG,Miller DR

    更新日期:2010-02-23 00:00:00

  • Ribosome recycling step in yeast cytoplasmic protein synthesis is catalyzed by eEF3 and ATP.

    abstract::After each round of protein biosynthesis, the posttermination complex (PoTC) consisting of a ribosome, mRNA, and tRNA must be disassembled into its components for a new round of translation. Here, we show that a Saccharomyces cerevisiae model PoTC was disassembled by ATP and eukaryotic elongation factor 3 (eEF3). GTP ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1006247107

    authors: Kurata S,Nielsen KH,Mitchell SF,Lorsch JR,Kaji A,Kaji H

    更新日期:2010-06-15 00:00:00

  • Glutamic and aminoadipic semialdehydes are the main carbonyl products of metal-catalyzed oxidation of proteins.

    abstract::Metal-catalyzed oxidation results in loss of function and structural alteration of proteins. The oxidative process affects a variety of side amino acid groups, some of which are converted to carbonyl compounds. Spectrophotometric measurement of these moieties, after their reaction with 2,4-dinitrophenylhydrazine, is a...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.011526698

    authors: Requena JR,Chao CC,Levine RL,Stadtman ER

    更新日期:2001-01-02 00:00:00

  • A point mutation in the regulatory light chain reduces the step size of skeletal muscle myosin.

    abstract::Current evidence favors the theory that, when the globular motor domain of myosin attaches to actin, the light chain binding domain or "lever arm" rotates, and thereby generates movement of actin filaments. Myosin is uniquely designed for such a role in that a long alpha-helix (approximately 9 nm) extending from the C...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0401699101

    authors: Sherwood JJ,Waller GS,Warshaw DM,Lowey S

    更新日期:2004-07-27 00:00:00

  • Notch signaling and the determination of appendage identity.

    abstract::The Notch signaling pathway defines an evolutionarily conserved cell-cell interaction mechanism that throughout development controls the ability of precursor cells to respond to developmental signals. Here we show that Notch signaling regulates the expression of the master control genes eyeless, vestigial, and Distal-...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.040556497

    authors: Kurata S,Go MJ,Artavanis-Tsakonas S,Gehring WJ

    更新日期:2000-02-29 00:00:00

  • Fluorescent metallacycle-cored polymers via covalent linkage and their use as contrast agents for cell imaging.

    abstract::The covalent linkage of supramolecular monomers provides a powerful strategy for constructing dynamic polymeric materials whose properties can be readily tuned either by the selection of monomers or the choice of functional linkers. In this strategy, the stabilities of the supramolecular monomers and the reactions use...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1612898113

    authors: Zhang M,Li S,Yan X,Zhou Z,Saha ML,Wang YC,Stang PJ

    更新日期:2016-10-04 00:00:00

  • Mg2+ mediates interaction between the voltage sensor and cytosolic domain to activate BK channels.

    abstract::The voltage-sensor domain (VSD) of voltage-dependent ion channels and enzymes is critical for cellular responses to membrane potential. The VSD can also be regulated by interaction with intracellular proteins and ligands, but how this occurs is poorly understood. Here, we show that the VSD of the BK-type K(+) channel ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0705873104

    authors: Yang H,Hu L,Shi J,Delaloye K,Horrigan FT,Cui J

    更新日期:2007-11-13 00:00:00

  • Growth hormone-releasing hormone antagonist MZ-5-156 inhibits growth of DU-145 human androgen-independent prostate carcinoma in nude mice and suppresses the levels and mRNA expression of insulin-like growth factor II in tumors.

    abstract::Insulin-like growth factors I and II (IGF-I and -II) are potent mitogens for various cancers, including carcinoma of the prostate. In several experimental cancers, treatment with antagonists of growth hormone-releasing hormone (GH-RH) produces a reduction in IGF-I and -II, concomitant to inhibition of tumor growth. To...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.15.8864

    authors: Lamharzi N,Schally AV,Koppán M,Groot K

    更新日期:1998-07-21 00:00:00

  • Actin antibody: the specific visualization of actin filaments in non-muscle cells.

    abstract::Actin purified from mouse fibroblasts by sodium dodecyl sulfate gel electrophoresis was used as antigen to obtain an antibody in rabbits. The elicited antibody was shown to be specific for actin as judged by immunodiffusion and complement fixation against partially purified mouse fibroblast actin and highly purified c...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.71.6.2268

    authors: Lazarides E,Weber K

    更新日期:1974-06-01 00:00:00

  • Solution structure of the silkworm betaGRP/GNBP3 N-terminal domain reveals the mechanism for beta-1,3-glucan-specific recognition.

    abstract::The beta-1,3-glucan recognition protein (betaGRP)/Gram-negative bacteria-binding protein 3 (GNBP3) is a crucial pattern-recognition receptor that specifically binds beta-1,3-glucan, a component of fungal cell walls. It evokes innate immunity against fungi through activation of the prophenoloxidase (proPO) cascade and ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0901671106

    authors: Takahasi K,Ochiai M,Horiuchi M,Kumeta H,Ogura K,Ashida M,Inagaki F

    更新日期:2009-07-14 00:00:00

  • Genetically modified anthrax lethal toxin safely delivers whole HIV protein antigens into the cytosol to induce T cell immunity.

    abstract::Bacillus anthrax lethal toxin can be engineered to deliver foreign proteins to the cytosol for antigen presentation to CD8 T cells. Vaccination with modified toxins carrying 8-9 amino acid peptide epitopes induces protective immunity in mice. To evaluate whether large protein antigens can be used with this system, rec...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.97.14.8027

    authors: Lu Y,Friedman R,Kushner N,Doling A,Thomas L,Touzjian N,Starnbach M,Lieberman J

    更新日期:2000-07-05 00:00:00

  • 3.3 Å structure of Niemann-Pick C1 protein reveals insights into the function of the C-terminal luminal domain in cholesterol transport.

    abstract::Niemann-Pick C1 (NPC1) and NPC2 proteins are indispensable for the export of LDL-derived cholesterol from late endosomes. Mutations in these proteins result in Niemann-Pick type C disease, a lysosomal storage disease. Despite recent reports of the NPC1 structure depicting its overall architecture, the function of its ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1711716114

    authors: Li X,Lu F,Trinh MN,Schmiege P,Seemann J,Wang J,Blobel G

    更新日期:2017-08-22 00:00:00

  • Binding of actinomycin D to DNA: evidence for a nonclassical high-affinity binding mode that does not require GpC sites.

    abstract::We have employed a combination of temperature-dependent UV absorption spectroscopy, circular dichroism, and batch calorimetry to characterize the binding of actinomycin D to a series of oligomeric DNA duplexes. We find the duplex [d(CGTCGACG)]2 to be unique in its ability to bind actinomycin D strongly despite the abs...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.11.3968

    authors: Snyder JG,Hartman NG,D'Estantoit BL,Kennard O,Remeta DP,Breslauer KJ

    更新日期:1989-06-01 00:00:00

  • Regulation of metallothionein genes by heavy metals appears to be mediated by a zinc-sensitive inhibitor that interacts with a constitutively active transcription factor, MTF-1.

    abstract::A construct, MRE-beta Geo, with five metal response elements fused to a selectable reporter gene was transfected into BHK cells and a stable clone that could be induced up to 100-fold by zinc, cadmium, bismuth, silver, cobalt, copper, mercury, or nickle was isolated. Some, and perhaps all, of these metals induce MRE-b...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.91.4.1219

    authors: Palmiter RD

    更新日期:1994-02-15 00:00:00

  • Transcription factor evolution in eukaryotes and the assembly of the regulatory toolkit in multicellular lineages.

    abstract::Transcription factors (TFs) are the main players in transcriptional regulation in eukaryotes. However, it remains unclear what role TFs played in the origin of all of the different eukaryotic multicellular lineages. In this paper, we explore how the origin of TF repertoires shaped eukaryotic evolution and, in particul...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1311818110

    authors: de Mendoza A,Sebé-Pedrós A,Šestak MS,Matejcic M,Torruella G,Domazet-Loso T,Ruiz-Trillo I

    更新日期:2013-12-10 00:00:00

  • Molecular mechanism for metal-independent production of hydroxyl radicals by hydrogen peroxide and halogenated quinones.

    abstract::We have shown previously that hydroxyl radicals (HO*) can be produced by H2O2 and halogenated quinones, independent of transition metal ions; however, the underlying molecular mechanism is still unclear. In the present study, using the electron spin resonance secondary radical spin-trapping method, we found that tetra...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0704030104

    authors: Zhu BZ,Kalyanaraman B,Jiang GB

    更新日期:2007-11-06 00:00:00

  • Dual-modality gene reporter for in vivo imaging.

    abstract::The ability to track cells and their patterns of gene expression in living organisms can increase our understanding of tissue development and disease. Gene reporters for bioluminescence, fluorescence, radionuclide, and magnetic resonance imaging (MRI) have been described but these suffer variously from limited depth p...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1319000111

    authors: Patrick PS,Hammersley J,Loizou L,Kettunen MI,Rodrigues TB,Hu DE,Tee SS,Hesketh R,Lyons SK,Soloviev D,Lewis DY,Aime S,Fulton SM,Brindle KM

    更新日期:2014-01-07 00:00:00

  • Structural basis for Zika envelope domain III recognition by a germline version of a recurrent neutralizing antibody.

    abstract::Recent epidemics demonstrate the global threat of Zika virus (ZIKV), a flavivirus transmitted by mosquitoes. Although infection is usually asymptomatic or mild, newborns of infected mothers can display severe symptoms, including neurodevelopmental abnormalities and microcephaly. Given the large-scale spread, symptom s...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1919269117

    authors: Esswein SR,Gristick HB,Jurado A,Peace A,Keeffe JR,Lee YE,Voll AV,Saeed M,Nussenzweig MC,Rice CM,Robbiani DF,MacDonald MR,Bjorkman PJ

    更新日期:2020-05-05 00:00:00

  • Optimizing transmembrane domain helicity accelerates insulin receptor internalization and lateral mobility.

    abstract::Transmembrane (TM) domains of integral membrane proteins are generally thought to be helical. However, a Gly-Pro sequence within the TM domain of the insulin receptor is predicted to act as a helix breaker. CD analyses of model TM peptides in a lipid-like environment show that substitution of Gly and Pro by Ala enhanc...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.12.5762

    authors: Goncalves E,Yamada K,Thatte HS,Backer JM,Golan DE,Kahn CR,Shoelson SE

    更新日期:1993-06-15 00:00:00

  • Evolutionary changes of multiple visual pigment genes in the complete genome of Pacific bluefin tuna.

    abstract::Tunas are migratory fishes in offshore habitats and top predators with unique features. Despite their ecological importance and high market values, the open-ocean lifestyle of tuna, in which effective sensing systems such as color vision are required for capture of prey, has been poorly understood. To elucidate the ge...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1302051110

    authors: Nakamura Y,Mori K,Saitoh K,Oshima K,Mekuchi M,Sugaya T,Shigenobu Y,Ojima N,Muta S,Fujiwara A,Yasuike M,Oohara I,Hirakawa H,Chowdhury VS,Kobayashi T,Nakajima K,Sano M,Wada T,Tashiro K,Ikeo K,Hattori M,Kuhara S,

    更新日期:2013-07-02 00:00:00

  • Genetic evidence that Acanthamoeba myosin I is a true myosin.

    abstract::Acanthamoeba castellanii contains two enzymes, myosins IA and IB, that exhibit the catalytic properties of a myosin but possess very unusual physical properties, the most striking of which are their single, low molecular weight heavy chain, their globular shape, and their inability to form bipolar filaments. We have n...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.83.13.4655

    authors: Hammer JA 3rd,Jung G,Korn ED

    更新日期:1986-07-01 00:00:00