EECS 1022 Chapter Notes - Chapter 2.6: Chromatin Immunoprecipitation, Gluconeogenesis, Glucocorticoid Receptor

• School: York University
• Department: Electrical Engineering and Computer Science
• Course: EECS 1022
• Professor: Cummins

Document Summary

After 6-8hr, the chimeric receptors will express a protein in the reporter cell line. Add drug candidates (possible ligands) and if ligand binds lbd, luciferase enzyme (reporter protein) will be generated and produce light in presence of substrate. Chipseq (chromatin immunoprecipitation sequencing) is used to identify nr binding sites on dna. Use formaldehyde to crosslink proteins to dna and shear dna into smaller soluble fragments. Add specific antibody to the protein of interest to make it precipitate and isolate it. Reverse cross-link the protein from dna using high heat or high salinity. Purify & sequence the dna to find the protein binding sites. 3 ways in which antagonists can prevent nr signalling: Failure to dimerize (due to bulky side chains of antagonist) Failure to recruit cofactors (due to bulky side chains of antagonist) 5 a. ) unliganded car in nucleus is active ligand shuts down response. Unliganded car in cytoplasm is inactive ligand activates response.