Abstract: A split luciferase-based sensor system was developed to noninvasively monitor and image phosphorylation-mediated c-Myc activation, in which the complementation of the split FL is induced by phosphorylation-mediated interaction between GSK3? and c-Myc. The complemented luciferase activity resulting from this interaction is specific to c-Myc phosphorylation and correlated with the steady-state and temporal regulation of c-Myc phosphorylation in cell culture. The sensor system also allows monitoring of c-Myc—targeted drug efficacy in intact cells and living animals. This new imaging sensor can provide insight into the role of functional c-Myc in cancer biology and is useful for the discovery and development of specific anti-c-Myc drugs.

Abstract: A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

Abstract: The present invention relates to the field of DNA recombinant technology. More specifically, this invention relates to fusion proteins comprising an ATP generating polypeptide joined to a polypeptide that converts ATP into a detectable entity. Accordingly, this invention focuses on sulfurylase-luciferase fusion proteins. This invention also relates to pharmaceutical compositions containing the fusion proteins and methods for using them.

Abstract: The present invention is concerned with test systems for determining the activity of neurotoxin polypeptides. Specifically, it relates to a polynucleotide encoding a single chain luciferase fusion polypeptide comprising: (i) a LuxB subunit, (ii) a linker comprising a neurotoxin cleavage site, and (iii) a LuxA subunit and a polypeptide encoded by the polynucleotide. Further provided in accordance with the invention are a vector and a host cell comprising the polynucleotide. Moreover, the present invention relates to a method for determining a proteolytically active neurotoxin polypeptide in a sample and a kit for carrying out the method.

Abstract: A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

Abstract: There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.

Abstract: The invention provides proliferative response indicator cell having a vertebrate cell having a luciferase encoding nucleic acid and a heterologous proliferation factor receptor encoding nucleic acid, wherein each of the encoding nucleic acids are operationally linked to expression elements for co-expression of a luciferase polypeptide and a heterologous proliferation factor receptor. The invention also provides a method of determining a cell proliferative response to a proliferation factor.

Abstract: The NF-E2-related factor 2 (Nrf2) is a key transcriptional regulator of antioxidant defense and detoxification. To directly monitor stabilization of Nrf2 we fused its Neh2 domain, responsible for the interaction with its nucleocytoplasmic regulator, Keap1, to firefly luciferase (Neh2-luciferase). It is shown herein that Neh2 domain is sufficient for recognition, ubiquitination and proteasomal degradation of Neh2-luciferase fusion protein. The novel Neh2-luc reporter system allows direct monitoring of the adaptive response to redox stress and classification of drugs based on the time-course of reporter activation. The novel reporter was used to screen a library of compounds to identify activators of Nrf2. The most robust and yet non toxic Nrf2 activators found—nordihydroguaiaretic acid, fisetin, and gedunin-induced astrocyte-dependent neuroprotection from oxidative stress via an Nrf2-dependent mechanism.

Abstract: The invention relates to compositions comprising a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to compositions comprising one or more polynucleotides encoding a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to methods and kits for detecting protein-protein interactions, determining the location of a protein-protein interaction, identifying cells wherein there is a protein-protein interaction of interest, and screening for a candidate modulator that increases or decreases the amount of a protein-protein interaction.

Abstract: A method, called GETWISE, for targeting mouse genes is described. GETWISE is designed to increase the frequency of homologous recombination, facilitate screening, widen the applicability of engineered animals and circumvent intrinsic gene targeting problems. GETWISE utilizes the principle of modulating gene expression by targeting tetracycline-responsive elements into a specific locus. In GETWISE alleles, control of gene expression is transferred from the endogenous to a tetracycline-inducible promoter. Endogenous promoters now control expression of the reporter gene luciferase. Breeding of GETWISE carriers with tTA/rtTA carriers enables investigators to modulate gene expression in a ubiquitous or tissue-specific manner, depending on the presence of doxycycline. GETWISE enables the study of loss or gain of gene expression in any tissue of choice within a single mouse strain. GETWISE enables the analysis of the gene expression pattern with the luciferase assay.

Abstract: The present invention provides nucleic acid constructs that encode fusion peptides comprising a bioluminescent protein and a precursor of a secreted peptide or protein expressed at the cell surface and high throughput screening assays using same.

Abstract: A gene construct incorporating any of at least two luciferase genes which emit lights with different colors using an identical substrate such that the gene can be stably expressed in mammalian cells.

Abstract: A tetracycline-regulated system which provides autoregulatory, inducible gene expression in cultured cells and transgenic animals is described. In the autoregulatory plasmid pTet-tTAk, a modified tTA gene called tTAk was placed under the control of Tetp. Tetracycline prevents tTA from binding to Tetp, preventing expression of both tTA and luciferase. This negative feedback cycle ensures that little or no tTA is produced in the presence of tetracycline, thereby reducing or eliminating possible toxic effects. When tetracycline is removed, however, this strategy predicts that tiny amounts of tTA protein (which may result from the leakiness of the minimal promoter), will bind to Tet-op and stimulate expression of the tTAk gene. A positive feedforward loop is initiated which in turn leads to higher levels of expression of tTA and thus, luciferase.

Abstract: The present invention provides methods and compositions useful in the diagnosis and management of autoimmune diseases. In particular, the present invention provides improved methods and compositions for the diagnosis and management of Graves' disease. The methods of the present invention not only avoids the need for radioactivity and are much simpler, economical, and rapid than methods traditionally used for the diagnosis of Graves' disease, but also improve upon the sensitivity and detection abilities of previous luciferase-based autoantibody detection assays.

Abstract: This invention is directed to a bioassay for determining the functionality of parathyroid hormone compounds. More particularly, this invention is directed to a bioassay wherein the compound to be tested is added to a culture of parathyroid hormone receptor expressing cells bearing a reporter gene under the transcriptional control of multiple c-AMP responsive elements.

Abstract: Isolated and purified nucleic acids encoding green fluorescent proteins from Renilla reniformis and the green fluorescent protein encoded thereby are also provided. Mutants of the nucleic acid molecules and the modified encoded proteins are also provided. Compositions and combinations comprising the green fluorescent proteins and/or the luciferase are further provided.

Abstract: The present invention provides methods and compositions useful in the diagnosis and management of autoimmune diseases. In particular, the present invention provides improved methods and compositions for the diagnosis and management of Graves' disease. The methods of the present invention not only avoids the need for radioactivity and are much simpler, economical, and rapid than methods traditionally used for the diagnosis of Graves' disease, but also improve upon the sensitivity and detection abilities of previous luciferase-based autoantibody detection assays.

Abstract: The present invention provides methods and compositions useful in the diagnosis and management of autoimmune diseases. In particular, the present invention provides improved methods and compositions for the diagnosis and management of Graves' disease. The methods of the present invention not only avoids the need for radioactivity and are much simpler, economical, and rapid than methods traditionally used for the diagnosis of Graves' disease, but also improve upon the sensitivity and detection abilities of previous luciferase-based autoantibody detection assays.

Abstract: The invention relates to methods and compositions which utilize the emission of light to monitor changes in microenvironments involving cells. The invention is especially useful for monitoring exocytotic activity such as detecting quantal release of synaptic vesicles. Fusion proteins of Cypridina luciferase and synaptotagmin-I or VAMP/synaptobrevin-2 were targeted to synaptic vesicles and, upon exocytosis, formed light-emitting complexes with luciferin present in the extracellular medium. Photon emissions in the presence of a depolarizing stimulus can be observed with these systems. pH-sensitive mutants of green fluorescent protein are also provided, which are useful for visualizing exocytosis and for imaging and measuring the pH of intracellular compartments.

Abstract: The embodiments of the present disclosure encompass methods for non-invasive in vivo bioluminescence imaging that allow the dynamics of stem cell behavior to be followed in a manner not possible using conventional retrospective static histological analyses. By imaging luciferase-generated bioluminescence activity emanating from isolated stem cells, for example, real time quantitative and kinetic analyses can show that donor-derived muscle stem cells may proliferate and engraft rapidly after injection until homeostasis is reached. In addition, the response of the stem cells to injury and participation in the regenerative response can be monitored over time. Other aspects of the disclosure encompasses methods for determining the suitability of a stem cell for tissue replacement, methods for repairing muscle injury, and methods for isolating muscle stem cells from a tissue sample.