Provided herein are chemically modified luciferase substrates for spectrally shifted emission and enhanced water solubility. Provided herein are engineered luciferases. Moreover, provided herein are new ATP-independent bioluminescent reporters which have improved biochemical and photophysical properties and are expected to have broad applications. Finally, provided herein are spectral-resolved triple-color bioluminescent systems, suitable for flexible and convenient approaches to monitor multiple biological events in either qualitative or quantitative manners

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 invention provides a method of designing an optimized gene which comprises altering a nucleotide sequence of a target protein gene, so that only preferential codons with high frequency of use in human cells are selected and a GC content of not less than 60% is achieved. A gene design method which involves the feature “only preferential codons with high frequency of use are selected and a GC content of not less than 60% is achieved” can be established as a general rule for preparing proteins with high expression level, in order to obtain chemically synthesized genes for proteins capable of high-level expression in eukaryotes.

Provided herein are bioluminescent polypeptides and compositions and methods for the assembly of a tripartite or multipartite bioluminescent complex. In particular embodiments, a bioluminescent complex is formed upon the interaction of three or more peptide and/or polypeptide components.

The present invention relates to bioluminescence resonance energy transfer sensor molecules having the structure R.sup.1-L-R.sup.2B or BR.sup.2-L-R.sup.1, wherein R.sup.1 is a bioluminescent protein, L is a linking element, R.sup.2 is a non-protein acceptor domain and B is a blocking group, and wherein R.sup.2 bound to B comprises a hydrolysable bond which produces a change in BRET when hydrolysed. The invention also discloses a method of detecting a hydrolase by contacting a sample with a molecule BR.sup.2, then contacting with a compound R.sup.1-L or L-R.sup.1 under conditions to cause attaching of R.sup.2 to L, and detecting a change in the BRET ratio. Specifically exemplified sensors comprise luciferase and fluorescein diacetate, which is hydrolysed by an esterase. The invention also discloses luciferase enzymes derived from RLuc8 by removing cysteine residues.

This disclosure relates to recombinant proteins, vectors, and methods of treating neurological conditions by exposing neurons to an opsin and luciferase in the presence of a luciferin. In certain embodiments, the disclosure relates to treating or preventing epilepsy or seizures comprising administering an effective amount of a vector that encodes an opsin and luciferase in combination with a luciferin to a subject in need thereof.

An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

In order to provide modified luciferase whose substrate specificity to at least one luminescent substrate (e.g., AkaLumine) other than D-luciferin has been improved as compared with to D-luciferin, modified luciferase according to an aspect of the present invention has a mutation at an amino acid corresponding to a 347th amino acid in an amino acid sequence represented by SEQ ID NO: 1.

An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

Provided herein are compositions and methods for the assembly of a tripartite or multipartite bioluminescent complex. In particular, a bioluminescent complex is formed upon the interaction of two or more peptide tags (e.g., separately or fused as a dipeptide or tripeptide) and a polypeptide component.