Resonance energy transfer (RET)- or protein-fragment complement assay (PCA)-based biosensors useful for assessing the activity of G-proteins are described. These biosensors are based on the competition between the Got subunit and a Gβγ interacting protein (βγ IP) for the binding to the Gβγ dimer. These biosensors comprises (1) a βγ IP and (2) a Gβ or Gγ protein; a GPCR; or a plasma membrane targeting domain, fused to suitable RET or PCA tags. Methods using such biosensors for different applications, including the identification of agents that modulates G-protein activity or for the characterization of GPCR signaling/regulation, such as G-protein preferences and activation profiles of GPCRs, are also described.

Disclosed herein are neuronal cell activity reporter systems including a Secreted Neuronal Activity Reporter (SNAR) construct and a control construct. The SNAR construct includes four tandem repeats of a core domain of the Synaptic Activity Response Element (SARE) of Arc/Arg3.1, a polynucleotide comprising the Arc minimal promoter, and a polynucleotide encoding a first secreted reporter protein. The control construct includes a constitutive promoter and a polynucleotide encoding a second secreted reporter protein. Further provided are methods of monitoring neuronal activity in a cell. The methods may include administering to a cell the neuronal cell activity reporter system, contacting with a substrate, and measuring a signal.

The present invention encompasses modified luciferases, methods for making modified luciferases, and assays utilizing modified luciferases. Modified luciferases of the invention show increased activity over wildtype luciferases and also show increased stability of signal. The present invention also encompasses multiplex assays utilizing multiple luciferases with different emission spectra.

Provided herein are isolated polynucleotide encoding modified click beetle luciferase polypeptides that have enhanced luminescence and longer wavelength near-infrared signals. The disclosure also relates to near-infrared bioluminescence systems that include said modified click beetle luciferase polypeptides and novel luciferin derivatives, as well as methods of using said modified click beetle luciferase polypeptides and bioluminescence systems.

Disclosed herein are methods, compositions, apparatus, systems and kits for performing polynucleotide amplification utilizing a pure polynucleotide polymerase. In some cases, disclosed herein are methods, compositions, apparatus, systems and kits for sequencing polynucleotides.

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.

According to aspects illustrated herein, there is provided an agent that transiently disrupts claudin-1 within tight junctions. The agent includes a peptide having at least 40% polar, uncharged amino acid residues and a self-assembled β-sheet secondary structure. According to aspects illustrated herein, there is also provided a transepithelial drug and vaccine formulations, as well as isolated peptides, pharmaceutical compositions, and transdermal delivery devices. Also described herein are methods of disrupting epithelial barrier and methods of administering the transepithelial formulations described herein.

Provided herein are substantially non-luminescent peptide/polypeptide tags that are inserted internally within a protein of interest or between N-terminal and C-terminal peptides/polypeptides. Interaction of the internally-inserted tag with a complement polypeptide/peptide that is also substantially non-luminescent results in the formation a bioluminescent reporter complex.

Efficient sequence specific gene silencing is possible through the use of siRNA technology. By selecting particular siRNAs by rational design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes. Methods, compositions, and kits generated through rational design of siRNAs are disclosed including those directed to nucleotide sequences for TTR.

Provided are a recombinant polynucleotide encoding a polypeptide including a reporter moiety, a substrate moiety, and a destabilization moiety, a host cell including the same, and use thereof to measure the level of a protease by using the recombinant polynucleotide.