Development of epigenetic CRISPR/Cas9 system for targeted methylation at specific CpG sites

Dobrinić, Paula (2016) Development of epigenetic CRISPR/Cas9 system for targeted methylation at specific CpG sites. Doctoral thesis, Faculty of Science > Department of Biology.

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Abstract

DNA methylation is an important epigenetic mechanism involved in genome regulation and normal development in mammals. CpG methylation of regulatory regions is usually associated with gene silencing, but exact mechanisms underlying this correlation are still unknown. Precise functional studies have been limited due to the lack of methods for targeted manipulation of methylation marks. To that end, I developed a flexible CRISPR/Cas9-based tool for targeted methylation of specific CpG sites. The programmable DNA binding module and DNA methyltransferase activity of DNMT3A were combined into a single fusion protein. Significant hypermethylation of targeted CpG sites at promoters of candidate genes was induced in human cultured cells, which resulted in gene transcriptional silencing. The established methylation profile was dependent on the associated complementary guide RNA. The described epigenetic tool could be very useful for precise interrogation of the causal relationship between DNA methylation and gene regulation.

Item Type: Thesis (Doctoral thesis)
Keywords: DNA methylation, epigenetic regulation, CRISPR/Cas9 system, epigenetic editing, gene silencing
Supervisor: Zoldoš, Vlatka
Date: 2016
Number of Pages: 81
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 28 Sep 2016 11:02
Last Modified: 28 Sep 2016 11:02
URI: http://digre.pmf.unizg.hr/id/eprint/5121

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