Methods of assaying receptor activity and constructs useful in such methods

• US 6,770,449 B2
• Filed: 01/30/2001
• Issued: 08/03/2004
• Est. Priority Date: 06/05/1997
• Status: Expired due to Term

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Zhang, Jie, Caron, Marc G., Barak, Lawrence S., Ferguson, Stephen S.
• Primary Examiner(s): FALK, ANNE MARIE
• Application Number:
  US09/772,644
• Publication Number:
  US 20020053093A1
• Time in Patent Office: 1,281 Days
• Field of Search: 435/7.2, 435/4, 435/350, 530/350
• US Class Current: 435/7.2
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0356 Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/60 containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

Described are methods of detecting G-protein coupled receptor (GPCR) activity in vivo and in vitro; methods of assaying GPCR activity; and methods of screening for GPCR ligands, G protein-coupled receptor kinase (GRK) activity, and compounds that interact with components of the GPCR regulatory process. Constructs useful in such methods are described.

Claims

1. A substrate having deposited thereon a plurality of cells, said cells expressing at least one GPCR and further comprising a biologically active labeled arrestin protein and wherein the label is capable of indicating localization of the arrestin and the GPCR is capable of binding the arrestin.

2. The substrate having deposited thereon a plurality of cells of claim 1, wherein the label is green fluorescent protein, β

-galactosidase, or luciferase.