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RADICL-ChIP: a Method to Capture Global RNA-on-Genome Distribution Mediated by Chromatin associated Factors

Chromatin associated RNAs play a key role in mediating epigenetic mechanism. To improve our comprehension of how chromatin-associated RNAs influence gene expression, it is crucial to identify the genomic locus that RNA interacts with on a genome-wide scale. Emerging technologies were developed to capture binding sites of lncRNAs globally. Such techniques rely on the concept of Proximity ligation via a biotinylated adapter to ligate DNA and RNA on each end, such as MARGI, GRID-seq, ChAR-seq and the most recent technology, devised in our lab RNA And DNA Complexes Ligated & Sequenced (RADICL-seq). RADICL-seq has several advantages over the other methods. However, while this method produced a fairly global picture of the chromatin associated “RNA-ome”, the specific association with specific regulatory factors is missing, in addition all emerging technologies lack the sensitivity to capture low-expressed RNAs. Thus, we set out a strategy to address this issue by enriching global RNA-chromatin interactions mediated by a specific factor which will be achieved by combining RADICL with Chromatin Immunoprecipitation (ChIP) method. Since our lab interest is investigating the role of lncRNAs in muscle differentiation, RADICL-ChIP has been performed using the C2C12 mouse skeletal muscle. In particular, the role of identification of RNA moieties interacting with PRC2 PcG proteins in stress response and their genome wide distribution in chromatin.

Identiferoai:union.ndltd.org:kaust.edu.sa/oai:repository.kaust.edu.sa:10754/673879
Date11 1900
CreatorsSaferuddin, Somiya
ContributorsOrlando, Valerio, Biological and Environmental Science and Engineering (BESE) Division, Li, Mo, Hirt, Heribert
Source SetsKing Abdullah University of Science and Technology
LanguageEnglish
Detected LanguageEnglish
TypeThesis
Rights2022-12-31, At the time of archiving, the student author of this thesis opted to temporarily restrict access to it. The full text of this thesis will become available to the public after the expiration of the embargo on 2022-12-31.

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