ISOLATION OF FACTORS THAT ASSOCIATE DIRECTLY OR INDIRECTLY WITH NON-CODING RNAS

摘要

Methods and assays are provided for isolating factors including polypeptides, ribonucleic acids (RNAs) and polypeptide complexes that are associated with a target nucleic acid sequence. The target nucleic acid sequence may be comprised within chromatin. The methods are suitable for identification and characterisation of factors including non-coding RNAs (ncRNAs) that associate with specified genomic loci.

主权项

1. A method for identifying one or more factors associated with a target nucleic acid sequence, wherein the one or more factors comprise at least one ribonucleic acid (RNA) sequence that is associated with the target nucleic acid sequence, the method comprising the steps of:
(a) obtaining a sample that comprises the target nucleic acid sequence and the one or more factors associated with the target nucleic acid sequence; (b) contacting the sample with one or more capture probes, wherein the capture probes comprise a nucleic acid sequence and at least one affinity label, and wherein the capture probes specifically hybridise with the at least one RNA sequence; (c) providing conditions that allow the one or more capture probes to hybridise with the at least one RNA sequence so as to form a hybridization complex between the capture probe, the at least one RNA, the target nucleic acid sequence and the one or more factors associated with the target nucleic acid sequence; (d) isolating the hybridization complex by immobilising the hybridization complex via a molecule that interacts with the affinity label; and (e) analyzing the constituents of the isolated hybridization complex so as to identify the one or more factors associated with the target nucleic acid sequence.