Your browser does not support JavaScript!

Post-graduate theses

Current Record: 24 of 800

Back to Results Previous page
Next page
Add to Basket
[Add to Basket]
Identifier 000460509
Title The role of the nascent RNA in chromatin conformation and genome regulation
Alternative Title Ο ρόλος του νεοσυντιθέμενου RNA στη ρύθμιση της διαμόρφωσης της χρωματίνης και της γονιδιακής έκφρασης
Author Κυδωνάκης, Κωνσταντίνος Ε.
Thesis advisor Lavigne, Matthieu
Reviewer Σπηλιανάκης, Χαράλαμπος
Γιακουντής, Αντώνιος
Abstract Nascent RNA has for long time been considered as a passive molecule of transcription and co-transcriptional processes. However, recently published studies demonstrate and establish its role in transcription progression, splicing and chromatin architecture. In line with these studies, we tried to standardize and optimize two molecular methods to address the issue of RNA organization. Our aim was to elaborate on its implication in various nuclear function, such as 3D chromatin conformation maintenance and gene expression regulation. We apply Global RNA Interaction with DNA by deep sequencing (GRID-seq) is an “all to all” genomic approach to study RNA-chromatin interaction and CRISPR-dCas9 system-based chromatin precipitation (CRISPR pull down) as a molecular approach to targeting specific chromatin regions and co-precipitate every factor that interacts with it. Regarding GRID-seq, we managed to perform double fixation in our cells, but further optimization is necessary, particularly in improving the efficiency of nuclei isolation. In parallel, for CRISPR pull down, we worked with the in vitro approach, and attempted to isolate the target chromatin and evaluate if our assay was specific calculating fold enrichment ratio between expected vs non-specific loci via qPCR. Although we only reached low efficiency in our first attempts, we propose several optimizations that will improve our approach.
Language English
Issue date 2023-11-24
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
Permanent Link https://elocus.lib.uoc.gr//dlib/6/6/a/metadata-dlib-1700653279-903492-11457.tkl Bookmark and Share
Views 962

Digital Documents
No preview available

Download document
View document
Views : 3