Technologies Available
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ACAT-2 Background: Cholesterol esterification in mammals is catalyzed by the intracellular enzyme acyl CoA:cholesterol acyltransferase (ACAT). In addition to its role in cellular cholesterol homeostasis, ACAT is important in several physiologic processes including intestinal cholesterol absorption, hepatic lipoprotein production, and the development of atherosclerotic lesions. Mouse tissue expression studies and the disruption of the mouse ACAT gene have indicated that more than one ACAT exists in mammals and, specifically, that another cholesterol acyltransferase enzyme is evident in mouse liver and intestine. Description: Researchers at the Gladstone Institutes have recently cloned the mouse and human cDNAs for the second ACAT enzyme, ACAT-2, and have identified that ACAT-2 mRNA is found primarily in mouse liver and intestine. This finding suggests that ACAT-2 is responsible for cholesterol esterification in these tissues. ACAT-2 exhibits a high specific activity with cholesterol as well as other oxysterol substrates, whereas ACAT-1 appears to be more active with oxysterols than with cholesterol. In addition, ACAT-2 exhibits different sensitivities from ACAT-1 towards various inhibitors. Applications: The availability of ACAT-2 reagents would facilitate the development of specific ACAT inhibitors that selectively target different forms of ACAT, for example, macrophage-derived ACAT-1, which is involved in atherosclerosis, and ACAT-2, which may be important in cholesterol absorption or lipoprotein assembly. Advantages: The Gladstone ACAT-2 gene would allow in vivo studies, such as gene disruption, which could elucidate the role of ACAT-2 in mammalian biology. Such studies were of major importance in defining the physiological role of ACAT-1. Reference: GL2006-80C See: U.S. Patent Nos. 6,579,974, 6,869,937 and 7,238,779 Gladstone Contact: Sandra Hall |
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ACAT-2 DEFICIENT MICE Background: Acyl CoA:cholesterol acyltransferase (ACAT) catalyzes the formation of cholesterol esters from cholesterol and fatty acids. Cholesterol esterification and the ACAT enzyme have been implicated in a number of important biological processes including cholesterol absorption, lipoprotein (such as LDL) synthesis and secretion, and macrophage foam cell formation in atherosclerotic lesions. Description: Scientists at the Gladstone Institutes have created a genetically modified strain of mice carrying a disruption of the mouse ACAT gene. This modification prevents these mice from making the ACAT enzyme. These mice exhibit markedly diminished cholesterol esterification in the adrenal cortical cells and in macrophages. Furthermore, the ACAT-deficient mice offer research advantages over treating mice with pharmaceutical ACAT inhibitors since the ACAT deficiency is complete and specific, avoiding any potential side effects of pharmaceutical agents. Applications: The Gladstone mice provide a model for examining the role of the ACAT enzyme in macrophage cholesterol ester foam cell formation and in atherosclerosis. Advantages: The ACAT-deficient mice permit investigations of the effects of blocking the enzymatic action of ACAT without the side effects of pharmaceutical agents. Reference: GL2006-80C See: U.S. Patent No. 6,579,974 Gladstone Contact: Sandra Hall |
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CALBINDIN-D28K: A RELIABLE INDICATOR AND POSSIBLE MEDIATOR OF NEURONAL DEFICITS IN ALZHEIMER'S DISEASE Background: Alzheimer's disease (AD) is a progressive neurodegenerative disease that causes a relentless decline in memory and other cognitive functions. It affects nearly half of those over 85, one in ten over 65, and even a small percentage of people in their 30s and 40s. The need for drugs that prevent or reverse the progression of AD in the long term is striking. Description: Since AD is a cognitive disorder, behavioral deficits in AD-related animal models are often regarded as the most relevant end point measures for the preclinical screening of novel drugs for this disease. However, behavioral testing is cumbersome, expensive, and time consuming. It often also gives inconsistent results when carried out in different laboratories. Using a well-characterized AD mouse model, UC researchers have discovered a strong correlation between behavioral deficits and loss of the calcium-binding protein, Calbindin-D28K (CB) in granules of the dentate gyrus in the brain. As opposed to performing the difficult and unreliable tests that are used to determine behavioral deficits directly, it is simple to reproducibly immunostain brain sections for CB. Since CB levels accurately reflect AD-related behavioral deficits, immunostaining for CB would allow for the testing of significantly larger numbers of drugs in larger numbers of mice at greater speed and reduced cost. In addition to simply performing as an indicator, CB-related pathways might also prove to play a causal role in the development of AD-related cognitive deficits. In this case, these findings could lead to the identification of novel drug targets for the treatment of this dreaded disease. Applications:
Reference: GL2006-815 See: U.S. Patent No. 7,297,836 Gladstone Contact: Sandra Hall |
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CELL LINES CARRYING LATENT HIV Background: Ongoing combination antiretroviral therapy can control the replication of human immunodeficiency virus (HIV) and delay the progression of AIDS. However, despite the complete suppression of detectable viremia in many patients during treatment, viremia reemerges rapidly after interruption of treatment, consistent with the existence of latent viral reservoirs. The presence of latent reservoirs has prevented the eradication of HIV from infected patients successfully treated with antiretroviral therapy. Knowledge of the mechanism by which the latent state is established, which is essential to its elimination, is poorly understood, partly because of the lack of an in vitro model. Prior to this, all stable cell lines in which HIV was latent carried a defective HIV. Description: Gladstone researchers have created model stable cell lines that harbor a latent, non-defective HIV provirus linked to a readily selectable, green fluorescent protein marker. The latency of infection can be broken and the virus activated by several known categories of chemicals. Purposeful activation and elimination of the proviruses that remain latent following treatment with combination antiretroviral therapy in patients would be one more giant step down the road to curing the scourge of AIDS. Applications:
Reference: GL2006-814 See: U.S. Patent Nos. 7,232,685 and 7,544,467 Gladstone Contact: Sandra Hall |
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CELL-SURFACE RECEPTOR FOR EBOLA AND MARBURG VIRUSES Background: Filoviruses such as the Marburg and Ebola viruses cause sporadic epidemics of human disease characterized by systemic hemorrhage, multi-organ failure and death in most instances. There is currently no accepted vaccine or direct therapy for the clinical manifestations of infection. Mechanisms by which these viruses gain entry into cells to initiate the replication cycle are poorly understood. Description: Gladstone researchers have cloned a cDNA encoding a receptor that has structural and functional characteristics that are consistent with its role in cellular entry by the Marburg and Ebola viruses. This receptor confers permissivity for infection by both Marburg and Ebola viruses to cells that were not originally capable of being infected. The identification of this cell surface mediator of filovirus entry offers direct evidence that Marburg and Ebola viruses share a common pathway for infection and may aid in the development of new therapeutic approaches. Applications:
Reference: GL2006-830 See: U.S. Patent Nos. 6,933,108 and 7,803,555 Gladstone Contact: Sandra Hall |
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CELL-BASED FUSION ASSAY FOR ANTIRETROVIRALS Background: Infection of mammalian cells by enveloped viruses typically involves proteinprotein interactions between glycoproteins expressed on the surface of the virion and specific "receptors" or cofactors expressed on the surface of the target cell. These interactions trigger fusion between the cellular and viral membranes in a process referred to as "entry." Description: It is recognized that this process can be mimicked by cellcell fusions in which the viral glycoproteins are expressed on the surface of a mammalian cell (representing the virion) and the host factors are expressed on the surface of a second mammalian cell (representing the target cell). Applications: Scientists at Gladstone have developed an assay system based on these cellcell fusion principles that is optimized for pharmaceutical screening purposes. In this assay, mixing two stable cell transfectants under defined conditions yields cellcell fusion and a fluorescence-based signal that can be easily quantitated using commerically available reagents and equipment. This invention, which can be readily adapted for other enveloped viruses, should prove useful to facilitate pharmaceutical discoveries. Reference: GL2006-810 See: U.S. Patent No. 6,451,598 Gladstone Contact: Sandra Hall |
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DGAT-1 Reference: GL2006-80D See: U.S. Patent No. 6,344,548 Gladstone Contact: Sandra Hall |
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DGAT-2 Reference: GL2006-812 See: U.S. Patent No. 7,045,326 Gladstone Contact: Sandra Hall |
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DGAT-1 KNOCKOUT MICE Reference: 1998-240 Gladstone Contact: Sandra Hall |
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DGAT-1 POLYMORPHISM ASSOCIATED WITH OBESITY Background: Gladstone scientists discovered polymorphisms in the human DGAT1 gene, a gene that encodes an enzyme involved in the synthesis of fat (triglycerol). One of the polymorphisms, a common variation in the promoter region of the human DGAT1 gene, has been correlated with body mass index, blood pressure, and HDL cholesterol levels in Turkish women. This common variation in the DNA may be useful in screening for propensity to conditions such as obesity or diabetes. Reference: GL2006-832 Patent Status: U.S. Patent Nos. 6,444,427 and 6,911,539 Gladstone Contact: Sandra Hall |
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HIGH-THROUGHPUT SCREENS FOR G PROTEIN-COUPLED RECEPTORS Background: G protein–coupled receptors (GPCRs) communicate extracellular signals to intracellular G proteins that transduce and amplify the signals. In mammals, it is believed that there are over 1000 GPCRs. Of those, several hundred are likely to be involved with various disease processes and thus are potential diagnostic and/or therapeutic targets. The majority of GPCRs activate G proteins that modulate cyclic AMP pathways, which unfortunately are not particularly amenable to large-scale screening methods. Furthermore, while the detection of calcium released by the phospholipase C pathway activated by certain G proteins is amenable to high-throughput or robotic screening methods, the GPCRs that activate these G proteins are relatively scarce. Description: Researchers at the Gladstone Institutes have created several G protein constructs that activate the phospholipase C pathway in response to various GPCRs that normally activate other pathways, such as the cAMP pathway. This provides a rapid means (less than 60 seconds) for identifying modulators of an increased number and variety of GPCRs by detecting the increased calcium levels using high-throughput screening methods. Examples of this technology are presented in J. Biol. Chem. 272:19107, 1997. Applications: Among the physiological responses involving signal transduction via GPCRs are the dilation/constriction of blood vessels, bronchi and organs within the gastrointestinal tract, the modulation of endocrine secretions, and the control of heart rate. This technology accelerates the screening for modulators (including agonists and antagonists) of GPCRs and, thus, has tremendous potential for the development of new diagnostics and therapeutics. Advantages: The Gladstone technology provides a rapid method (less than 60 seconds) to identify modulators of an increased number and variety of GPCRs. The method is:
Reference: GL2006-826 See: U.S. Patent No. 6,383,761 Gladstone Contact: Sandra Hall |
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HIGHLY SENSITIVE TEST FOR DETECTION OF ANTIGEN-SPECIFIC T CELL RESPONSES Background: Exposure to HIV, other viral and some bacterial and parasite diseases is detected by assays that measure immune responses in vitro. The enzyme-linked immunosorbent spot assay (ELISPOT) assay has become widely used as a tool for the detection of these cellular and humoral immune responses. Although the sensitivity and technical ease of the ELISPOT assay make it a useful alternative to traditional analytical methods, there remains a need for increased sensitivity to detect low level antigen-specific T cell responses in patients that have been difficult to diagnose by standard methods. The addition of autologous dendritic cells has been shown to augment the sensitivity of ELISPOT assays. However, this method requires large amounts of fresh peripheral blood and in vitro maturation of monocytes over a period of 5 to 7 days. It is therefore desirable to identify a combination of cytokines and/or co-stimulatory molecules that are able to substitute for the potent antigen-presenting capability of mature dendritic cells. Description: Researchers at the University of California have discovered that addition of the gamma-c cytokines IL-7 and IL-15 significantly enhances the sensitivity of the ELISPOT assay. Sensitivity in fresh samples was augmented up to 7.5-fold, and, using cryopreserved specimens, sensitivity was augmented up to 18-fold. Applications: Increasing the sensitivity of the ELISPOT assay will prove useful for:
Advantages:
Reference: GL2006-854 See: U.S. Patent No. 7,364,869 Gladstone Contact: Sandra Hall |
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HUMAN HDAC7 Background: Acetylation of core histones has been correlated with transcription, chromatin assembly, DNA repair, and recombinational events. Transfer of an acetyl group from acetyl–CoA onto the epsilon–amino group of different lysine residues in the NH2–terminal tail of core histones is a ubiquitous modification found in all euykaryotic species examined. Histone acetylation levels are controlled by the competing activities of histone acetyltransferases and histone deacetylases. Histone deacetylases are the catalytic subunits of multiprotein complexes that are targeted to specific promoters through their interaction with sequence–specific DNA–binding factors. Description: Gladstone scientists have identified, cloned and characterized a novel human histone deacetylase (HDAC). This protein is highly expressed in specific populations of T lymphocytes in the thymus and at lower levels in other tissues. This newly discovered histone deacetylase is associated with histone deacetylase activity that is dependent on binding another identified histone deacetylase. Applications: Since HDACs are involved in the control of gene regulation, HDAC genes are potential candidates as tumor suppressors. As such, their mutation could be associated with the development of cancers such as lymphomas, thymomas, or tumors of other organs. A probe of the newly discovered HDAC, or probes derived from its sequence, could become important tools for the diagnosis of specific cancers. Similarly, preparation and use of specific inhibitors of Gladstone’s newly discovered HDAC could play a significant role in the control of immunoproliferation, such as in immunosuppression for organ transplantation, as well as in the control of tumor development. Reference: GL2006-836 See: U.S. Patent No. 7,488,587 Gladstone Contact: Sandra Hall |
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HUMAN SIRT3 Background: The yeast silent information regulator (Sir)2 protein links cellular metabolism and transcriptional silencing through its nicotinamide adenine dinucleotide (NAD)–dependent histone deacetylase activity. Description: Gladstone scientists discovered that mitochondria from mammalian cells contain intrinsic NAD–dependent deacetylase activity. This activity is inhibited by the NAD hydrolysis product nicotinamide, but not by trichostatin A, consistent with a class III deacetylase. Gladstone scientists identified this deacetylase as the nuclear–encoded human Sir2 homologue hSIRT3, and show that hSIRT3 is located within the mitochondrial matrix. Mitochondrial import of hSIRT3 is dependent on an NH2–terminal amphipathic–helix rich in basic residues. hSIRT3 is proteolytically processed in the mitochondrial matrix to a 28-kD product. Applications: This processing can be reconstituted in vitro with recombinant mitochondrial matrix processing peptidase (MPP) and is inhibited by mutation of arginines 99 and 100. The unprocessed form of hSIRT3 is enzymatically inactive and becomes fully activated in vitro after cleavage by MPP. These observations demonstrate the existence of a latent class III deacetylase that becomes catalytically activated upon import into the human mitochondria. Reference: 2002-350 See: U.S. Patent No. 7,273,713 Gladstone Contact: Sandra Hall |
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INHIBITOR OF TNF–α AND IL-1 Background: Stimulation of cells with various pro-inflammatory cytokines such as TNF–a and IL–1 begins a complex signaling pathway culminating in the translocation of a transcription factor, NF–kB, to the nucleus. NF–kB controls genes involved in inflammatory, apoptotic and immune responses. Description: Gladstone scientists have developed a novel inhibitor of the inflammatory response induced by TNF–a and IL–1. The inhibitor specifically prevents the phosphorylation, and thus activation, of the kinases IKKa and IKKb. These kinases play a critical role in the signaling cascade that leads to the translocation of NF–kB to the nucleus. Applications: Inhibition of phosphorylation of IKKa and IKKb is an efficient and specific way to block the activation of NF–kB following cytokine stimulation. The inhibitor can be used therapeutically to block inflammatory responses induced by TNF–a and IL–1. Reference: GL2006-828 See: U.S. Patent Nos. 6,265,538 and 6,645,728 Sandra Hall |
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METHOD TO IDENTIFY EDITED mRNAs Background: Messenger RNAs can be modified post–transcriptionally, by a process known as RNA editing, to produce a protein product or products different from what would be predicted from the genomic DNA sequence. There are several enzymes involved in editing mRNA. Overexpression of one such RNA editing protein in the liver and intestine of transgenic mice leads to the development of liver cancer or obesity, implying that RNA editing normally plays a critical role in controlling these diseases. Description: Gladstone scientists have developed a method to identify and clone edited genes from a library of cDNAs or in electronic databases of sequenced DNA. Application: The new method can be used to identify the edited genes involved in cancer and obesity. More generally, the methods can be used to identify and clone any novel edited gene. Reference: GL2006-824 See: U.S. Patent No. 5,866,333 Gladstone Contact: Sandra Hall |
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MGAT-1, MGAT-2, AND RELATED GENES Reference: GL2006-812 See: U.S. Patent No. 7,045,326 Gladstone Contact: Sandra Hall |
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MONOCLONAL ANTIBODY AGAINST HUNTINGTIN PROTEIN Background: Huntingtin’s disease (HD) is a progressive neurodegenerative condition with an autosomal dominant inheritance pattern. The gene responsible for the disorder encodes a protein known as huntingtin. Towards the 5’ end of the gene is a variable number of the glutamine-encoding nucleotide triplet CAG. The pathological version of the huntingtin protein contains an expanded tract of greater than 35 glutamines. Description: Gladstone scientists have raised a highly specific mouse monoclonal antibody against the long glutamine stretch in the pathological version of the huntingtin protein. The researchers have also isolated the gene encoding this antibody. Applications: The antibody could be used therapeutically to disrupt pathological interactions with the mutant huntingtin protein. It could also be used in high throughput screens to detect drugs that bind huntingtin and/or interfere with aggregation or other potentially pathological protein–protein interactions. Reference: GL2006-80A See: U.S. Patent No. 6,291,652 Gladstone Contact: Sandra Hall |
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NUCLEIC ACIDS ENCODING DOMINANT POSITIVE IκBα POLYPEPTIDES Background: Nuclear factor kB (NF–kB) is a eukaryotic transcription factor that exerts pleiotropic effects on diverse cellular genes involved in the immediate early steps of immune activation and inflammation. Additionally, NF–kB has been implicated in the transcriptional activation of several viruses, including HIV–1 (for a review, see Siebenlist et al. Annu. Rev. Cell. Biol. 10:405-455 (1994)). Nuclear expression and consequent biological action of the eukaryotic NF–kB transcription factor complex is tightly regulated through its cytoplasmic retention by an ankyrin-rich inhibitory protein termed IkBa. IkBa specifically binds to and masks the nuclear localization signal of the Rel A subunit of NF–kB, thereby effectively sequestering this transcription factor complex in the cytoplasm. Specific cellular activation signals lead to the rapid proteolytic degradation of IkBa and the concomitant nuclear translocation of NF–kB Such signals include, for example, mitogens such as phorbol esters, cytokines such as tumor necrosis factor alpha (TNF–a) and interleukin-1 (IL-1), and the Tax protein from the type I human T cell leukemia virus (HTLV–1). Activation of NF–kB by these and other inducers appears to involve the transient phosphorylation and subsequent proteolytic degradation of IkBa which permits nuclear translocation of the liberated NF–kB complex. Nuclear expression of the NF–kB complex leads to transcriptional activation of a broad array of cellular genes involved in immune stimulation, inflammation, and cell growth. Description: Scientists from the Gladstone Institutes constructed a class of dominant positive IkBa mutants that retains full inhibitory function of NF–kB yet fails to undergo stimulus induced degradation. Applications: These mutants can be used to inhibit NF–kB activation in specific target cells, thereby reducing or eliminating undesirable consequences of NF–kB activation. One aspect of the invention is a method of inhibiting NF–kB activation in a cell comprising introducing into the cell nucleic acid encoding a dominant positive mutant IkBa polypeptide, wherein the IkBa mutant polypeptide is expressed in the cell. A further aspect of the invention is a composition comprising an IkBa polypeptide, wherein residues 1 through 36 and 278-317 are deleted. Reference: GL2006-804 See: U.S. Patent No. 6,180,772 Gladstone Contact: Sandra Hall |
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RECEPTORS ACTIVATED SOLELY BY A SYNTHETIC LIGAND Background: A critical limitation of current in vivo and ex vivo gene therapy efforts is the inability to amplify the number of transfected cells. DNA encoding the therapeutic protein of interest can be successfully delivered to and expressed in the target cells, but often too few target cells are transformed to provide a measurable therapeutic response. If these few transformed cells could be specifically activated at the cellular level in vivo, e.g., to proliferate or to secrete their therapeutic product, the desired therapeutic effect could be achieved. Description: Researchers at the Gladstone Institutes have developed a powerful and versatile new method for control of cell processes, including proliferation and product secretion, in both in vitro and in vivo environments. The technology allows selective activation of the G-protein signaling pathway in target cells that express a receptor activated superiorly by a synthetic ligand (RASSL). These receptors have been engineered to be unresponsive to their natural ligands but remain responsive to synthetic small-molecule drugs. In addition to proliferation and secretion, cell processes that could be controlled by G-protein signaling pathway include cell migration, cell contraction, pigment production, heart rate control, models of human disease, and apoptosis, depending on the cell type expressing the RASSL. Additional information is available at Conklin Lab Homepage. Applications:
Advantages:
Reference: GL2006-823 See: U.S. Patent No. 6,518,480 Gladstone Contact: Sandra Hall |
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REGULATION OF NFκB Background: NFkB, a heterodimer of the proteins p50 and RelA, is an inducible eykaryotic transcription factor complex that plays a pivotal role in regulating both the inflammatory and immune responses in mammalian cells. By means of binding to regulatory regions of cellular targets, NFkB increases transcription of genes that function in inflammatory, immune and anti-apoptotic responses. Activation of NFkB is implicated in a variety of chronic inflammatory diseases. In addition, the replication of many viruses including HIV is enhanced by the activation of NFkB. NFkB in unstimulated cells is sequestered in the cytoplasm, and thus tightly regulated, by its assembly with the inhibitory protein IkBa. Description: Researchers at the Gladstone Institute of Virology and Immunology have discovered that RelA, the transcriptionally active component of the NFkB heterodimer, is acetylated in vivo. Such acetylated forms of RelA bind more effectively to DNA but fail to interact with IkBa. RelA is subsequently deacetylated by histone deacetylase 3 (HDAC3). This deacetylation reaction promotes IkBa binding and nuclear export of the NFkB complex. Acordingly, the NFkB transcriptional response is terminated and the latent cytoplasmic pool of NFkB is replenished. Reversible acetylation of RelA thus forms an intra-nuclear molecular switch for regulating NFkB action. These findings for the first time link deacetylation to the nuclear export of a eukaryotic transcription factor. In addition, RelA is identified as a novel nonhistone substrate regulated (deacetylated) by HDAC3. Applications: This discovery will allow screening assays to identify agents that are useful as therapeutics for modulating NFkB transcriptional activity. These approaches could have significant implication for the development of new classes of drugs exhibiting anti-neoplastic, anti-inflammatory, antiviral or immunoregulatory activity. Regulation of NFkB activity through the use of agents that block acetylation or increase deacetylation could provide a fundamentally new approach to inhibiting NFkB action. Agents that decrease the level of NF-kB-mediated transcription are desirable to reduce inflammation and increase apoptosis, among other effects. Agents that increase the levels of NFkB may be desirable to treat various forms of immunodeficiency. Reference:GL2006-835 See: U.S. Patent No. 7,081,343 Gladstone Contact: Sandra Hall |
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ROBOTIC MICROSCOPE Background: A new invention, a robotic microscope, is opening the way for scientists to track changes in cells over time. Images collected by the microscope can automatically be analyzed within minutes by a computer program. For example, a user can ask the computer to measure cells with a specific morphology, amounts of proteins, or other features. The robotic microscope system is the brainchild of an Investigator of the Gladstone Institute of Neurological Disease. Applications: The robotic microscope facilitates high-throughput cell biology. In a typical experiment, 300,000 cells are analyzed, a task that used to take 6 weeks. With the robotic microscope, it takes only 15 minutes. Another advantage of the new device is that the criteria used to define features of interest, such as markers of degeneration, are explicilty defined and uniformly applied by the computer program, eliminating user bias. The microscope can also be used to determine the rate of cell loss over time. Reference:GL2006-813 See: U.S. Patent 7,139,415 Gladstone Contact: Sandra Hall |
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SELECTION OF RNA-BINDING PEPTIDES FROM COMBINATORIAL LIBRARIES Description: Scientists at the Gladstone Institutes have designed a method to screen peptide libraries in order to identify peptides that bind a specific RNA sequence. The method employs an in vivo screen based on transcriptional antitermination in bacteria. The screening procedure can use relatively small combinatorial libraries constructed with restricted sets of amino acids to select RNA–binding peptides that, in principle, should bind any desired RNA target with high specificity and affinity. In verifying this system, the Gladstone scientists used their method to find peptides that bind the Rev–response element (RRE) of HIV. Not only did the screen pull out peptides that match the naturally occurring Rev consensus sequence, but it also pulled out peptides that are quite different from Rev, yet still bind the RRE RNA very tightly and specifically. Applications: This Gladstone screen has many possible uses in medicine and biotechnology. The ability to select molecules that bind to specific nucleic acid sites not only provides information about nucleic acid recognition, but also provides new strategies for altering gene expression and designing new therapeutics. The identification of sequence–specific RNA–binding peptides has great potential in the design of inhibitors, activators, and diagnostic markers of the expression of particular genes. Advantages:
Reference: GL2006-843 See: U.S. Patent No. 5,834,184 Gladstone Contact: Sandra Hall |
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TRANSGENIC RATS AND RAT CELL LINES EXPRESSING HUMAN CD4 AND A HUMAN CHEMOKINE RECEPTOR Background: The entry of HIV virus into the target ìhelperî T-cells and other target cells requires the interaction of HIV with cell-surface CD4 molecules and a chemokine receptor. Transgenic rabbits and mice models expressing human CD4 have been generated in an effort to develop transgenic animal models for the study of AIDS. However, differences in the infectivity between the rabbit and mouse models have been noticed due to species-specific cofactors that mediate post-entry viral processes. Although these animal models have been useful in research, there is a need for animal models that provide a more efficient and cost effective method of studying HIV infection in vivo and in vitro. Description: Researchers at The J. David Gladstone Institutes have developed a transgenic rodent animal model for the study of HIV infection and replication. The transgenic rodent is characterized by the expression of a human CD4 receptor and one or more human chemokine receptors. These receptors are stably integrated into the genome and enhance HIV infectivity, HIV structural genes expression, and HIV replication within the cells of the transgenic rodent, both in vivo and in vitro. Applications:
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Reference: GL2006-805 See: U.S. Patent No. 6,372,956 Gladstone Contact: Sandra Hall |
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TREATMENT AND ANIMAL MODEL FOR ANXIETY Background: Anxiety is a condition characterized by abnormally high levels of worry, uneasiness, apprehension and uncertainty. Mice lacking the ApoE gene exhibit symptoms of anxiety that can be measured by assessing their performance on standardized behavioral assays. Their anxiety can be rescued by transgenic addition of ApoE3 isotype, but not ApoE4. Description: Gladstone scientists have discovered agents that can be used to treat anxiety through the inhibition of ApoE4 or enhancement of ApoE3 activity. They have developed a transgenic mouse model and behavioral assessment techniques that can be used to identify and test anxiolytic drugs. Applications: The methods and drugs will be useful for treatment of anxiety. Reference: GL2006-831 See: U.S. Patent No. 6,982,361 Gladstone Contact: Sandra Hall |
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VIRION-BASED FUSION ASSAY Background: The entry of enveloped viruses into their host cells has been the focus of considerable attention, since this forms one of the earliest steps in the viral life cycle. Fusion mediated through viral envelopes to the cell plasma membrane has been studied in various cell–based fusion assays in which the cell expressing the viral envelope protein mimics the viral membrane. Although these techniques have been very useful in the past, with the recent advancement of cellular biology, it is becoming clear that the virion membrane differs significantly from the plasma membrane of the cell and that the nature of the target cell greatly influences the process of fusion. In addition, using cell–cell fusion assays it is difficult to study fusion in primary cells that form the biologically relevant targets in vivo. Two virion–based fusion assays have been developed so far, but their low sensitivity and high complexity have sharply limited their use. Description: Researchers at the Gladstone Institute of Virology and Immunology have developed a virion–based fusion assay that allows the detection of the fusion of HIV–1 (or HIV–1 pseudotyped viruses) to various target cells by microscopy, flow cytometry, or UV spectrometry. The assay incorporates an enzymatic amplification reaction that provides for high sensitivity, high specificity, and ease of use. The strategy is based on the delivery of the chimeric protein beta–lactamase/Vpr from the virus to the target cell as indicator of fusion. The transfer of this chimeric virion protein can be enzymatically revealed by monitoring the hydrolysis of a beta–lactamase fluorescent substrate pre–loaded in the target cells. This changes its fluorescence emission from green to blue, thereby providing an easy method of detection. Advantages:
Applications:
Reference: GL2006-838 See: U.S. Patent No. 7,250,251 Gladstone Contact: Sandra Hall |
Material Transfer Request
To initiate a new request for Gladstone research materials, please e-mail the appropriate investigator from this list of Gladstone investigators. Once approved, your request will be forwarded to our office for processing and you can expect to receive a material request form from our office soon thereafter.
