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Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy
dc.contributor | Dobrijević, Zorana | |
dc.creator | Sarić, Ana | |
dc.creator | Rajić, Jovana | |
dc.creator | Tolić, Anja | |
dc.creator | Đorđević, Marija | |
dc.creator | Grdović, Nevena | |
dc.creator | Uskoković, Aleksandra | |
dc.creator | Dinić, Svetlana | |
dc.creator | Arambašić Jovanović, Jelena | |
dc.creator | Mihailović, Mirjana | |
dc.creator | Vidaković, Melita | |
dc.creator | Dučić, Tanja | |
dc.date.accessioned | 2023-11-11T13:40:20Z | |
dc.date.available | 2023-11-11T13:40:20Z | |
dc.date.issued | 2023 | |
dc.identifier.issn | 2787-2947 | |
dc.identifier.uri | http://radar.ibiss.bg.ac.rs/handle/123456789/6304 | |
dc.description.abstract | Introduction: DNA methylation is a major regulator of transcriptional activity and alongside other epigenetic modifications it introduces specific level of chromatin complexity. Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive, and label-free technique for identifying subtle changes in all bio-macromolecules, and it has been used as a method of choice for studying DNA conformation. The present study was designed to explore the use of synchrotron-based FTIR spectroscopy to monitor the subtle changes on molecular level regarding the DNA methylation status of cytosine in the whole genome. Methods: For FTIR-based DNA methylation analysis in situ, DNA-HALO samples were prepared using slightly modified methodology for nuclear HALO preparations where DNA-HALOs are liberated of any protein residues but preserve higher order chromatin structure. Results: Using FTIR spectroscopy we analysed and compared methylation profiles of isolated genomic DNA and DNA-HALO samples. DNA-HALO structure shows more distinct peaks in fingerprint region of spectra. DNA-HALO structure is more accurate for detecting bonds in unmemthylated cytosine as specific infrared peaks are defined as vibrations of bonds in unmethylated cytosine at 1151 cm-1 and 1357 cm-1. The ratio of integrated area under the peak 1151 cm-1 over integrated area under the peak that represents PO2-backbone vibrations can be used to assess level of unmethylated cytosine and thus methylation rate in the DNA-HALO samples. Conclusion: This study demonstrates potential of FTIR spectroscopy to detect DNA methylation in DNA-HALO samples more precisely compared to classical DNA extraction procedure that yield unstructured whole genomic DNA. | sr |
dc.language.iso | en | sr |
dc.publisher | Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade | sr |
dc.relation | ALBA In-house grant: “Towards FTIR spectroscopy of single living cells” PI T. Ducic, 2018093034 | sr |
dc.relation | COST Action CA1406 | sr |
dc.relation | COST Action CA16119 | sr |
dc.relation | info:eu-repo/grantAgreement/MESTD/inst-2020/200007/RS// | sr |
dc.rights | openAccess | sr |
dc.source | Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia | sr |
dc.subject | HALO structure | sr |
dc.subject | FTIR spectroscopy | sr |
dc.subject | DNA methylation | sr |
dc.subject | Epigenetic marks | sr |
dc.subject | Chromatin | sr |
dc.title | Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy | sr |
dc.type | conferenceObject | sr |
dc.rights.license | ARR | sr |
dc.rights.holder | © 2023 by the Institute of Molecular Genetics and Genetic Engineering, University of Belgrade | sr |
dc.description.other | Dobrijević Z, editor. Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade ; 2023. p. 68. (Trends in Molecular Biology; Special Issue). | sr |
dc.citation.spage | 68 | |
dc.citation.epage | 68 | |
dc.type.version | publishedVersion | sr |
dc.identifier.fulltext | https://radar.ibiss.bg.ac.rs/bitstream/id/15457/bitstream_15457.pdf | |
dc.citation.rank | M64 | |
dc.identifier.rcub | https://hdl.handle.net/21.15107/rcub_ibiss_6304 |