SELEX against whole-cell bacteria resulted in lipopolysaccharide binding aptamers

dc.authorid0000-0002-1677-644X
dc.contributor.authorYılmaz, Deniz
dc.contributor.authorMuslu, Tuğdem
dc.contributor.authorParlar, Ayhan
dc.contributor.authorKurt, Hasan
dc.contributor.authorYüce, Meral
dc.date.accessioned2022-07-04T09:17:44Z
dc.date.available2022-07-04T09:17:44Z
dc.date.issued2022
dc.departmentİstanbul Medipol Üniversitesi, Mühendislik ve Doğa Bilimleri Fakültesi, Biyomedikal Mühendisliği Bölümü
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Sağlık Bilim ve Teknolojileri Araştırma Enstitüsü
dc.description.abstractNucleic acid aptamers are target-specific oligonucleotides selected from combinatorial libraries through an iterative in vitro screening process known as Systemic Evolution of Ligands by Exponential Enrichment (SELEX). In this report, the selection of bacteria differentiating ssDNA aptamer candidates from a combinatorial library through the whole-cell SELEX method was performed. The enriched SELEX pool was sequenced using Illumina Next-Generation Sequencing (NGS) technology and analyzed for the most abundant sequences using CLC Genomics Workbench. The sequencing data resulted in several oligonucleotide families from which three individual sequences were chosen per SELEX based on the copy numbers. The binding performance of the selected aptamers was assessed by flow cytometry and fluorescence spectroscopy, and the binding constants were estimated using binding saturation curves. Varying results were obtained from two independent SELEX procedures where the SELEX against the model gram-negative bacterium Escherichia coli provided more selective sequences while the SELEX library used against gram-positive bacterium Listeria monocytogenes did not evolve as expected. The sequences that emerged from E. coli SELEX were shown to bind Lipopolysaccharide residues (LPS) and inhibit LPS-induced macrophage polarization. Thus, it can be said that, performed whole-cell SELEX could be resulted as the selection of aptamers which can bind LPS and inhibit LPS induced inflammation response and thus can be candidates for the inhibition of bacterial infections. In future studies, the selected aptamer sequences could be structurally and chemically modified and exploited as potential diagnostic tools and therapeutic agents as LPS antagonists.
dc.identifier.citationYılmaz, D., Muslu, T., Parlar, A., Kurt, H. ve Yüce, M. (2022). SELEX against whole-cell bacteria resulted in lipopolysaccharide binding aptamers. Journal of Biotechnology, 354, 10-20. http://doi.org/10.1016/j.jbiotec.2022.06.001
dc.identifier.doi10.1016/j.jbiotec.2022.06.001
dc.identifier.endpage20
dc.identifier.issn0168-1656
dc.identifier.issn1873-4863
dc.identifier.pmid35700936
dc.identifier.scopus2-s2.0-85132529647
dc.identifier.scopusqualityQ2
dc.identifier.startpage10
dc.identifier.urihttp://doi.org/10.1016/j.jbiotec.2022.06.001
dc.identifier.urihttps://hdl.handle.net/20.500.12511/9558
dc.identifier.volume354
dc.identifier.wos000822628400002en_US
dc.identifier.wosqualityQ2
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorKurt, Hasan
dc.language.isoen
dc.publisherElsevier B.V.
dc.relation.ispartofJournal of Biotechnologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/embargoedAccess
dc.subjectAptamer
dc.subjectSELEX
dc.subjectNext Generation Sequencing
dc.subjectMacrophage
dc.subjectLPS
dc.titleSELEX against whole-cell bacteria resulted in lipopolysaccharide binding aptamers
dc.typeArticle

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