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CRISPR/dCas9 molecular tools reveal the regulation of FUT8, MGAT4A, MGAT4B, MGAT5, MGAT3, and B4GALT1 genes by CpG methylation (CROSBI ID 725399)

Prilog sa skupa u časopisu | sažetak izlaganja sa skupa

Vujić, Ana ; Klasić, Marija ; Vičić Bočkor, Vedrana ; Zoldoš, Vlatka ; Pučić Baković, Maja ; Lauc, Gordan CRISPR/dCas9 molecular tools reveal the regulation of FUT8, MGAT4A, MGAT4B, MGAT5, MGAT3, and B4GALT1 genes by CpG methylation // Journal of bioanthropology. 2022. str. 152-152 doi: 10.54062/jb

Podaci o odgovornosti

Vujić, Ana ; Klasić, Marija ; Vičić Bočkor, Vedrana ; Zoldoš, Vlatka ; Pučić Baković, Maja ; Lauc, Gordan

engleski

CRISPR/dCas9 molecular tools reveal the regulation of FUT8, MGAT4A, MGAT4B, MGAT5, MGAT3, and B4GALT1 genes by CpG methylation

In hepatocellular carcinoma (HCC), as well as in various other cancers, protein glycosylation is altered and as such is associated with tumor proliferation, invasion, metastasis, angiogenesis, and multidrug resistance. Mechanisms are mostly epigenetic. Indeed, aberrant DNA methylation is one of the epigenetic modifications that is highly perturbed in cancer, resulting in changes in transcriptional activity of many key genes, thus leading to characteristic cancer behavior. One group of genes, which might be affected, are glyco-genes coding for glycosyltransferases. The aim of this study is to explore epigenetic regulation of glyco-genes using cutting-edge CRISPR/Cas9 based molecular tools for epigenome editing. In hepatocellular carcinoma model cell line HepG2, we targeted seven candidate glyco-genes using dCas9-DNMT3A and dCas9-TET1 fusions, and subsequently analyzed CpG methylation, transcriptional gene activity, and whole-cell N-glycome as a final phenotype. Transfected cells were collected at two time points (8th and 12th day following transfection). Targeted methylation of selected CpG sites in ST6GAL1, FUT8, MGAT4A, MGAT4B, MGAT5, and B4GALT1 genes induced hypermethylation at these sites (up to 40% on average, depending on the gene), which was followed by a statistically significant change in transcriptional activity of all these genes except ST6GAL1. Targeted demethylation of MGAT3 gene (up to 45% on average) was accompanied by a statistically significant change in its transcription. As a final phenotype, whole-cell protein glycosylation was analyzed and changes in several glycosylation traits in HepG2 glycome were observed. These results suggest that alterations in CpG methylation lead to differential expression of glycosyltransferases, thus leading to aberrant protein glycosylation in HCC.

epigenome editing, gene regulation, CRISPR/dCas9, DNA methylation, protein glycosylation

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Podaci o prilogu

152-152.

2022.

nije evidentirano

objavljeno

10.54062/jb

Podaci o matičnoj publikaciji

Journal of bioanthropology

Zagreb: Institut za antropologiju

978-953-57695-4-5

2787-8201

Podaci o skupu

12th ISABS Conference on Forensic and Anthropologic Genetics and Mayo Clinic Lectures in Individidualized Medicine

poster

22.07.2022-27.07.2022

Dubrovnik, Hrvatska

Povezanost rada

Biologija

Poveznice