# US 11,579,149 B2 · Hippo pathway bioluminescent biosensor > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,579,149 B2 - Document ID: US11579149B2 - Status: Granted - Kind: B2 - Application number: 16/177,691 - Filed: 2018-11-01 - Published: 2019-08-22 - Granted: 2023-02-14 - Earliest claimed: 2017-11-01 - Assignees: Queen's University at Kingston - Applicants: Queen's University at Kingston - Inventors: Xiaolong Yang; Taha Azad; Kazem Nouri - Web page: https://patentgrep.com/patent/US11579149B2 ## CPC - G01N 33/582: {with fluorescent label} - C07K 14/4702: {Regulators; Modulating activity} - C12Q 1/66: involving luciferase - C07K 14/43563: {from insects} - C07K 2319/61: containing an enzyme fusion for detection (lacZ, luciferase) - C07K 2319/71: containing domain for transcriptional activaation, e.g. VP16 - C12N 9/0069: {acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)} - C12Y 113/12013: Oplophorus-luciferin 2-monooxygenase (1.13.12.13) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES ## Abstract Bioluminescent biosensors useful for monitoring and/or quantifying, in vitro or in vivo, activity of the Hippo signaling pathway. The biosensors monitor LATS kinase activity or YAP-TEAD interaction. The biosensors may be used in methods for monitoring and/or quantifying in real-time, in vitro or in vivo, activity of the Hippo signaling pathway, wherein the activity may be LATS kinase activity and/or YAP-TEAD interaction. The biosensors may be provided in kits for monitoring and/or quantifying in real-time, in vitro or in vivo, activity of the Hippo signaling pathway, wherein the activity may be LATS kinase activity and/or YAP-TEAD interaction. ## Claims ### Claim 1 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of firefly luciferase fused to at least one fragment of YAP. ### Claim 2 (depends on 1) The DNA construct of claim 1, wherein the fragment of firefly luciferase comprises amino acids 1-416 of SEQ ID NO:6 and is N-terminal to the at least one fragment of YAP comprising amino acids 120-134 of SEQ ID NO:2. ### Claim 3 (depends on 1) The DNA construct of claim 1, wherein the fragment of firefly luciferase comprises amino acids 1-544 of SEQ ID NO:6 and is N-terminal to the at least one fragment of YAP comprising amino acids 120-134 of SEQ ID NO:2. ### Claim 4 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of firefly luciferase fused to at least one fragment of 14-3-3 protein, wherein the at least one fragment of 14-3-3 protein is capable of binding to YAP. ### Claim 5 (depends on 4) The DNA construct of claim 4, wherein the at least one fragment of 14-3-3 protein comprises SEQ ID NO:4 and is N-terminal to the fragment of firefly luciferase comprising amino acids 394-550 of SEQ ID NO:6. ### Claim 6 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of NanoBiT luciferase fused to at least one fragment of YAP. ### Claim 7 (depends on 6) The DNA construct of claim 6, wherein the fragment of NanoBit luciferase is LgBiT comprising amino acid sequence SEQ ID NO:52 and is N-terminal to the at least one fragment of YAP comprising amino acids 120-134 of SEQ ID NO:2. ### Claim 8 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of NanoBiT luciferase fused to at least one fragment of 14-3-3 protein, wherein the at least one fragment of 14-3-3 protein is capable of binding to YAP. ### Claim 9 (depends on 8) The DNA construct of claim 8, wherein the at least one fragment of 14-3-3 protein comprises SEQ ID NO:4 and is N-terminal to the fragment of NanoBit luciferase that is SmBiT comprising amino acid sequence SEQ ID NO:54. ### Claim 10 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of NanoBit luciferase that is LgBiT comprising amino acid sequence SEQ ID NO:52 and a fragment of YAP comprising amino acids 50-171 of SEQ ID NO:2. ### Claim 11 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of NanoBit luciferase that is SmBiT comprising amino acid sequence SEQ ID NO:54 and a fragment of YAP comprising amino acids 50-171 of SEQ ID NO:2. ### Claim 12 A DNA construct comprising a nucleic acid sequence encoding a fusion protein comprising a fragment of NanoBiT luciferase fused to at least one fragment of TEAD protein, wherein the at least one fragment of TEAD protein is capable of binding to YAP. ### Claim 13 (depends on 12) The DNA construct of claim 12, wherein the fragment of NanoBit luciferase is LgBiT comprising amino acid sequence SEQ ID NO:52 and the at least one fragment of TEAD protein comprises amino acids 194-411 of SEQ ID NO:50. ### Claim 14 (depends on 12) The DNA construct of claim 12, wherein the fragment of NanoBit luciferase is SmBiT comprising amino acid sequence SEQ ID NO:54 and the at least one fragment of TEAD protein comprises amino acids 194-411 of SEQ ID NO:50.