RNA-YY1 interactions
Methods relating to obtaining libraries of YY1-binding long non-coding RNAs, libraries obtained thereby, and methods of use thereof.
1. A method of identifying a compound that disrupts binding of a Yin-Yang 1 (YY1)-binding long non-coding RNA (lncRNA) to Yin-Yang 1 (YY1) protein, the method comprising:
(a) identifying an lncRNA that binds YY1 protein;
(b) providing a cell-free sample comprising a YY1-binding lncRNA and YY1 protein, wherein the YY1-binding lncRNA binds to the YY1 protein thereby forming lncRNA-YY1 protein complexes;
(c) contacting the cell-free sample with a test compound, wherein the test compound is a nucleic acid targeting the YY1-binding lncRNA;
(d) isolating the lncRNA-YY1 protein complexes from the cell-free sample by contacting the cell-free sample comprising the lncRNA-YY1 protein complexes with an anti-YY1 antibody, wherein the anti-YY1 antibody specifically binds to YY1 protein, and immunoprecipitating the lncRNA-YY1 protein complexes; and
(e) detecting formation of the lncRNA-YY1 protein complexes in presence and absence of the test compound, wherein a decrease in formation of lncRNA-YY1 protein complexes in the presence of the test compound as compared to formation of lncRNA-YY1 protein complexes in the absence of test compound indicates that the test compound disrupts the binding of the YY1-binding lncRNA to the YY1 protein.
2. The method of claim 1 , wherein the YY1 protein is labeled.
3. The method of claim 1 , further comprising wherein the YY1 protein not bound to YY1-binding lncRNA is isolated from the cell-free sample.
4. The method of claim 1 , wherein the YY1-binding lncRNA is labeled.
5. The method of claim 1 , wherein both the YY1 protein and the YY1-binding lncRNA are labeled.