The instant disclosure relates to methods for luciferase modification.

The present invention relates to provision of novel artificial bioluminescent enzymes. According to one embodiment of the present invention, there is provided a polypeptide, including any one of amino acid sequences (i) to (iii) below, and having a copepod luciferase activity: (i) an amino acid sequence represented by SEQ ID NO: 1 or 12; (ii) an amino acid sequence represented by SEQ ID NO: 1 or 12 in which one or several amino acids are substituted, added, or deleted; or (iii) an amino acid sequence having an identity of not less than 90% with an amino acid sequence represented by SEQ ID NO: 1 or 12.

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.

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.

Described are coelenterazine analogs, methods for making the analogs, kits comprising the analogs, and methods of using the compounds for the detection of luminescence in luciferase-based assays.

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.

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.

The present invention provides, among other things, imidazo[1,2-?]pyrazine derivatives including ##STR00001##
The derivatives are useful in any method which other coelenterazines have been used. For example, the derivatives may be used in a bioluminogenic method to detect the presence of certain compounds or molecules.

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.

Described are coelenterazine analogues, methods for making the analogues, kits comprising the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.