A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.

Provided herein are compositions comprising a double-stranded RNA (dsRNA) binding domains linked to components of a complementation system. Upon binding of a pair of dsRNA binding domains to a dsRNA, a detectable complex of the complementation components is formed, and the dsRNA can be detected/quantified.

Cas-cleavable reporter systems, CRISPR-Cas systems thereof, and methods of use thereof in CRISPR-Cas based diagnostics. Cas-cleavable reporter systems may generate a luminescent, fluorescent, or other detectable signal upon Cas-collateral cleavage of one or more reporter system components. CRISPR-Cas systems may comprise a Cas protein having collateral cleavage activity, guide molecules, and the Cas-cleavable reporter system. Cas-cleavable reporter systems may be a split luciferase reporter system, at least one element of which is bound to beads. For multiplexing, CRISPR-Cas systems may comprise a Cas-cleavable quenched reporter system, optionally a Cas-cleavable quenched fluorescent reporter system, a Cas protein having collateral cleavage activity, and target-specific guide molecules attached to color-coded beads. CRISPR-Cas systems may further comprise amplification reagents. Methods may apply said CRISPR-Cas systems to flow cell, well-plate, or other devices, and may measure detectable signals by microscopic, plate reader, or other methods.

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.

Described herein is a generalizable strategy to rapidly and irreversibly activate protein function with full spatiotemporal control. Through development of an exogenously triggerable self-assembling protein construct, bioactive proteins can be stably reassembled from non-functional split fragment pairs following exposure to a stimulus (e.g., light).

The present disclosure is directed to RNA-regulated fusion proteins comprising a protein of interest and an RNA-regulated destabilization domain. Also disclosed are RNA aptamers that bind specifically to a RNA-regulated destabilization domain. Nucleic acid molecules encoding the RNA-regulated fusion proteins and RNA aptamers and methods of use thereof are also disclosed.

The present disclosure provides a novel lipid nanoparticle (LNP) compositions useful for the transfection and expression of exogenous proteins in platelet cells. In particular, wherein the LNP composition comprises mRNA encoding the exogenous protein and a lipid mixture, and the lipid mixture comprises at least one ionizable cationic lipid, at least one helper lipid, a sterol, and at least one polyethylene glycol (PEG)-lipid conjugate. Further provided are use of the LNP compositions for transfection of platelets with messenger RNA encoding an exogenous protein and expression of the exogenous protein in the resulting transfected platelets.

A bioluminescent single photon bioreactor for performing absolute quantification of light-producing activity by enzymes includes: a bioreactor that produces a bio-electronic signal; an electronic sensor that receives the bio-electronic signal and produces an electrical transduction signal; and an analyzer that receives the electrical transduction signal and absolutely quantifies light-producing activity by enzymes from the electrical transduction signal, such that the absolute quantification is accomplished quantum mechanically by determination of a second order autocorrelation function.

Disclosed are ATP-independent bioluminescent reporter systems for enhanced in vivo imaging. Also disclosed is a modified luciferin, potassiorin, comprising a potassium-binding moiety. Also disclosed is an engineered luciferase, BRIPO, optimized for synergistic interaction with potassiorin. A further version of the disclosed system produces bioluminescence signals responsive to physiological potassium concentrations. In further instances, the system enables real-time monitoring of K+ dynamics in live cells, tissues, and animals. A further embodiment includes applications in imaging neuronal activity, studying ion flux, and developing bioluminescent indicators for diverse analytes. The disclosed system addresses limitations of traditional imaging methods, offering improved sensitivity and biocompatibility.