IP Library › Granted Patent US 12,655,398
Granted Patent B2
US 12,655,398 · App. 16/439,565 · Granted Jun 16, 2026

Multipartite luciferase

Inventors: Mary Hall (Madison, WI); Lance P. Encell (Madison, WI); Michael Killoran (Madison, WI); Keith Wood (Madison, WI); Thomas Smith (Madison, WI); Virginia Kincaid (Madison, WI); Thomas Kirkland (Madison, WI); Melanie Dart (Madison, WI)
Assignee: Promega Corporation
C12N9/0069C12Y113/12G01N21/763G01N33/582G01N33/6845
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,655,398
App. No.
16/439,565
Granted
Jun 16, 2026
Kind
B2
Abstract

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.

Claims (12)

1 . A composition comprising:

(a) a peptide comprising an amino acid sequence having 100% sequence identity with SEQ ID NO: 268, wherein the amino acid sequence is compared to the full length of SEQ ID NO: 268, wherein a bioluminescent signal produced in the presence of a coelenterazine substrate is capable of substantial increase when the peptide contacts a second peptide consisting of SEQ ID NO: 25 and a polypeptide complement consisting of SEQ ID NO: 17 when compared to a bioluminescent signal produced by (1) the peptide and the coelenterazine substrate alone or (2) the peptide, the coelenterazine substrate, and only one of the second peptide or the polypeptide complement;

(b) a nucleic acid comprising a sequence coding for the peptide of (a);

(c) a fusion protein comprising the peptide of (a) and additional amino acid sequence; or

(d) a nucleic acid comprising a sequence coding for the fusion protein of (c).

2 . The composition of claim 1 , wherein the bioluminescent signal is substantially increased when the peptide associates with the second peptide and the polypeptide complement.

3 . The composition of claim 1 , wherein the peptide exhibits enhancement of one or more traits compared to a peptide of SEQ ID NO: 6 and/or SEQ ID NO: 9 , wherein the traits are selected from: affinity for the second peptide and the polypeptide complement, expression, solubility, stability, and bioluminescent activity when combined with the second peptide and the polypeptide complement.

4 . The composition of claim 1 , wherein the additional amino acid sequence of the fusion is selected from the group consisting of a protein of interest, an interaction element, a co-localization element, and a binding moiety.

5 . The composition of claim 4 , wherein the additional amino acid sequence of the fusion is a binding moiety selected from the group consisting of antibody, antibody fragment, protein A, an Ig binding domain of protein A, protein G, an Ig binding domain of protein G, protein A/G, an Ig binding domain of protein A/G, protein L, an Ig binding domain of protein L, protein M, an Ig binding domain of protein M, peptide nucleic acid, DARPin, aptamer, affimer, a purified protein, and analyte binding domains of proteins.

6 . The composition of claim 5 , wherein the additional amino acid sequence of the fusion is a first interaction polypeptide that is configured to form a complex with a second interaction polypeptide upon contact of the first interaction polypeptide and the second interaction polypeptide.

7 . The composition of claim 4 , wherein the additional amino acid sequence of the fusion is a first co-localization polypeptide that is configured to co-localize within a cellular compartment, a cell, a tissue, or an organism within a with a second co-localization polypeptide.

8 . The composition of claim 4 , wherein the additional amino acid sequence of the fusion is a protein of interest and is a candidate drug target.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 21, 2021
From: HALL, MARY; ENCELL, LANCE P; KILLORAN, MICHAEL; WOOD, KEITH; SMITH, THOMAS; KINCAID, VIRGINIA; KIRKLAND, THOMAS; DART, MELANIE
To: PROMEGA CORPORATION
Reel/Frame 056310/0479 →
Continuity (2)
Provisional Application 62684014 · Jun 12, 2018
Related Publication 20200270586A1 · Aug 27, 2020
References Cited (71)
US 4946778A · Ladner et al. · 1990 [cited by applicant]
US 5260203A · Ladner et al. · 1993 [cited by applicant]
US 8669103B2 · Binkowski et al. · 2014 [cited by applicant]
US 8697359B1 · Zhang · 2014 [cited by applicant]
US 9056885B2 · Kirkland et al. · 2015 [cited by applicant]
US 9797889B2 · Dixon et al. · 2017 [cited by applicant]
US 9797890B2 · Dixon et al. · 2017 [cited by applicant]
US 9908918B2 · Lin et al. · 2018 [cited by applicant]
US 20060194256A1 · Miao · 2006 [cited by examiner]
US 20100281552A1 · Encell et al. · 2010 [cited by applicant]
US 20120056073A1 · Ahn · 2012 [cited by applicant]
US 20120107849A1 · Klaubert et al. · 2012 [cited by applicant]
US 20120174242A1 · Binkowski et al. · 2012 [cited by applicant]
US 20140068797A1 · Doudna et al. · 2014 [cited by applicant]
US 20140099654A1 · Cali et al. · 2014 [cited by applicant]
US 20140348747A1 · Dixon et al. · 2014 [cited by applicant]
US 20150152395A1 · Branchini · 2015 [cited by applicant]
US 20170233789A1 · Shakhmin et al. · 2017 [cited by applicant]
US 20180030059A1 · Hall et al. · 2018 [cited by applicant]
US 20180172692A1 · Dixon · 2018 [cited by examiner]
US 20200333346A1 · Dixon · 2020 [cited by examiner]
WO WO8801649 · 1988 [cited by applicant]
WO WO9306868 · 1993 [cited by applicant]
WO WO9408629 · 1994 [cited by applicant]
WO WO9409056 · 1994 [cited by applicant]
WO WO9626754 · 1996 [cited by applicant]
WO WO2003040100 · 2003 [cited by applicant]
WO WO2009128056A2 · 2009 [cited by examiner]
WO WO2012061529 · 2012 [cited by applicant]
WO WO2014151736 · 2014 [cited by applicant]
WO WO2017189751 · 2017 [cited by applicant]
WO WO2017189751A1 · 2017 [cited by examiner]
Inouye et al 2014; Biochem & Biophys Res Commun: vol. 445; pp. 157-162. (Year: 2014). [cited by examiner]
Extended European Search Report for PCT/U.S. Pat. No. 2019036844. Mailed May 8, 2022. 10 pages. [cited by applicant]
Dec. 14, 2017. Split enzyme developing beta-barrel protein peptide beta 9, seq id:2. XP002807009, retrieved from EBI accession No. GSP:BEN35145 Data base accession No. BEN 35145. 1 page. [cited by applicant]
Dec. 14, 2017. Split enzyme developing luciferase third polypeptide, SEQ ID:3. XP002807010, retrieved from EBI accession No. GSP:BEN35146. Database accession No. BEN35146. 1 page. [cited by applicant]
Dec. 14, 2017 Target screening beta 10*-His Tag-heavy chain polypeptide, SEQ ID:22. XP002807011. Retrieved from EBI accession No. GSP:BEN35165. 1 page. [cited by applicant]
Jiang et al., Lighting up bioluminescence with coelenterazine: strategies and applications. Photochem Photobiol Sci. Apr. 2016;15(4):466-80. [cited by applicant]
Barrangou et al., CRISPR provides acquired resistance against viruses in prokaryotes. Science. Mar. 23, 2007;315(5819):1709-12. [cited by applicant]
Bhaya et al., CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation. Annu Rev Genet. 2011;45:273-97. [cited by applicant]
Bolotin et al., Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology (Reading). Aug. 2005;151(Pt 8):2551-2561. [cited by applicant]
Boutureira et al., Advances in chemical protein modification. Chem Rev. Mar. 11, 2015;115(5):2174-95. [cited by applicant]
Brouns et al., Small CRISPR RNAs guide antiviral defense in prokaryotes. Science. Aug. 15, 2008;321(5891):960-4. [cited by applicant]
Cong et al., Multiplex genome engineering using CRISPR/Cas systems. Science. Feb. 15, 2013;339(6121):819-23. [cited by applicant]
Deltcheva et al., CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature. Mar. 31, 2011;471(7340):602-7. [cited by applicant]
Xiao-Jie et al., CRISPR-Cas9: a new and promising player in gene therapy. J Med Genet. May 2015;52(5):289-96. [cited by applicant]
Deveau et al., CRISPR/Cas system and its role in phage-bacteria interactions. Annu Rev Microbiol. 2010;64:475-93. [cited by applicant]
Garneau et al., The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature. Nov. 4, 2010;468(7320):67-71. [cited by applicant]
Gasiunas et al, Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc Natl Acad Sci U S A. Sep. 25, 2012;109(39):E2579-86. [cited by applicant]
Hale et al., RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex. Cell. Nov. 25, 2009;139(5):945-56. [cited by applicant]
Horvath et al., CRISPR/Cas, the immune system of bacteria and archaea. Science. Jan. 8, 2010;327(5962):167-70. [cited by applicant]
Huang et al., Production of Gene-Corrected Adult Beta Globin Protein in Human Erythrocytes Differentiated from Patient iPSCs After Genome Editing of the Sickle Point Mutation. Stem Cells. May 2015;33(5):1470-9. [cited by applicant]
Hudson et al., Engineered antibodies. Nat Med. Jan. 2003;9(1):129-34. [cited by applicant]
Inagaki et al., Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging. Sci Rep. Feb. 13, 2017;7:42398. [cited by applicant]
Isidro-Llobet et al., Amino acid-protecting groups. Chem Rev. Jun. 2009;109(6):2455-504. [cited by applicant]
Jinek et al., A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. Aug. 17, 2012;337(6096):816-21. [cited by applicant]
Koniev et al., Developments and recent advancements in the field of endogenous amino acid selective bond forming reactions for bioconjugation. Chem Soc Rev. Aug. 7, 2015;44(15):5495-551. [cited by applicant]
Makarova et al., A putative RNA-interference-based immune system in prokaryotes: computational analysis of the predicted enzymatic machinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of act… [cited by applicant]
Makarova et al., Evolution and classification of the CRISPR-Cas systems. Nat Rev Microbiol. Jun. 2011;9(6):467-77. [cited by applicant]
Mali et al., RNA-guided human genome engineering via Cas9. Science. Feb. 15, 2013;339(6121):823-6. [cited by applicant]
Marraffini et al., CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science. Dec. 19, 2008;322(5909):1843-5. [cited by applicant]
Mojica et al., Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J Mol Evol. Feb. 2005;60(2):174-82. [cited by applicant]
Patterson et al., Finding the right (bioorthogonal) chemistry. ACS Chem Biol. Mar. 21, 2014;9(3):592-605. [cited by applicant]
Pourcel et al., CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology (Reading). Mar. 2005;151(Pt 3):653-… [cited by applicant]
Sapranauskas et al., The [cited by applicant]
Schaub et al., Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical Imaging and Flow Cytometry for Monitoring Tumorigenesis. Cancer Res. Dec. 1, 2015;75(23):5023-33. [cited by applicant]
Smith et al., Efficient and allele-specific genome editing of disease loci in human iPSCs. Mol Ther. Mar. 2015;23(3):570-7. [cited by applicant]
Suzuki et al., Five colour variants of bright luminescent protein for real-time multicolour bioimaging. Nat Commun. Dec. 14, 2016;7:13718. [cited by applicant]
Tomalia et al., Starburst Dendrimers: Molecular-level control of size, shape, surface chemistry, topology, and flexibility from atoms to macroscopic Matter. Angewandte Chemie International Edition in English, 1990, 29(2… [cited by applicant]
Xie et al., Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac. Genome Res. Sep. 2014;24(9):1526-33. [cited by applicant]
Office Action for Chinese Application No. 2019800531393, dated Jan. 30, 2024 (10 pages). [cited by applicant]