Modified RNA triplexes: Thermodynamics, structure and biological potential

Abstract The occurrence of triplexes in vivo has been well documented and is determined by the presence of long homopurine-homopyrimidine tracts. The formation of these structures is the result of conformational changes that occur in the duplex, which allow the binding of a third strand within the m...

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Autores principales:	Marta Szabat, Elzbieta Kierzek, Ryszard Kierzek
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c556e732ad6a44c9aecdd40fa9be3673
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spelling	oai:doaj.org-article:c556e732ad6a44c9aecdd40fa9be36732021-12-02T15:08:25ZModified RNA triplexes: Thermodynamics, structure and biological potential10.1038/s41598-018-31387-52045-2322https://doaj.org/article/c556e732ad6a44c9aecdd40fa9be36732018-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-31387-5https://doaj.org/toc/2045-2322Abstract The occurrence of triplexes in vivo has been well documented and is determined by the presence of long homopurine-homopyrimidine tracts. The formation of these structures is the result of conformational changes that occur in the duplex, which allow the binding of a third strand within the major groove of the helix. Formation of these noncanonical forms by introducing synthetic triplex-forming oligonucleotides (TFOs) into the cell may have applications in molecular biology, diagnostics and therapy. This study focused on the formation of RNA triplexes as well as their thermal stability and biological potential in the HeLa cell line. Thermodynamics studies revealed that the incorporation of multiple locked nucleic acid (LNA) and 2-thiouridine (2-thioU) residues increased the stability of RNA triplexes. These data suggest that the number and position of the modified nucleotides within TFOs significantly stabilize the formed structures. Moreover, specificity of the interactions between the modified TFOs and the RNA hairpin was characterized using electrophoretic mobility-shift assay (EMSA), and triplex dissociation constants have been also determined. Finally, through quantitative analysis of GFP expression, the triplex structures were shown to regulate GFP gene silencing. Together, our data provide a first glimpse into the thermodynamic, structural and biological properties of LNA- and 2-thioU modified RNA triplexes.Marta SzabatElzbieta KierzekRyszard KierzekNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Marta Szabat Elzbieta Kierzek Ryszard Kierzek Modified RNA triplexes: Thermodynamics, structure and biological potential
description	Abstract The occurrence of triplexes in vivo has been well documented and is determined by the presence of long homopurine-homopyrimidine tracts. The formation of these structures is the result of conformational changes that occur in the duplex, which allow the binding of a third strand within the major groove of the helix. Formation of these noncanonical forms by introducing synthetic triplex-forming oligonucleotides (TFOs) into the cell may have applications in molecular biology, diagnostics and therapy. This study focused on the formation of RNA triplexes as well as their thermal stability and biological potential in the HeLa cell line. Thermodynamics studies revealed that the incorporation of multiple locked nucleic acid (LNA) and 2-thiouridine (2-thioU) residues increased the stability of RNA triplexes. These data suggest that the number and position of the modified nucleotides within TFOs significantly stabilize the formed structures. Moreover, specificity of the interactions between the modified TFOs and the RNA hairpin was characterized using electrophoretic mobility-shift assay (EMSA), and triplex dissociation constants have been also determined. Finally, through quantitative analysis of GFP expression, the triplex structures were shown to regulate GFP gene silencing. Together, our data provide a first glimpse into the thermodynamic, structural and biological properties of LNA- and 2-thioU modified RNA triplexes.
format	article
author	Marta Szabat Elzbieta Kierzek Ryszard Kierzek
author_facet	Marta Szabat Elzbieta Kierzek Ryszard Kierzek
author_sort	Marta Szabat
title	Modified RNA triplexes: Thermodynamics, structure and biological potential
title_short	Modified RNA triplexes: Thermodynamics, structure and biological potential
title_full	Modified RNA triplexes: Thermodynamics, structure and biological potential
title_fullStr	Modified RNA triplexes: Thermodynamics, structure and biological potential
title_full_unstemmed	Modified RNA triplexes: Thermodynamics, structure and biological potential
title_sort	modified rna triplexes: thermodynamics, structure and biological potential
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/c556e732ad6a44c9aecdd40fa9be3673
work_keys_str_mv	AT martaszabat modifiedrnatriplexesthermodynamicsstructureandbiologicalpotential AT elzbietakierzek modifiedrnatriplexesthermodynamicsstructureandbiologicalpotential AT ryszardkierzek modifiedrnatriplexesthermodynamicsstructureandbiologicalpotential
_version_	1718388148069728256

Modified RNA triplexes: Thermodynamics, structure and biological potential

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