| アイテムタイプ |
リポジトリ登録用アイテムタイプ（シンプル）(1) |
| 公開日 |
2026-04-06 |
| タイトル |
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タイトル |
High-Throughput Sequencing and SELEX-Based Protocol for Selecting Aptamers Against Potato Spindle Tuber Viroid |
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言語 |
en |
| 言語 |
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言語 |
eng |
| 資源タイプ |
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資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
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資源タイプ |
journal article |
| アクセス権 |
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アクセス権 |
open access |
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アクセス権URI |
http://purl.org/coar/access_right/c_abf2 |
| 著者 |
Kaponi, Maria S.
| en |
Kaponi, Maria S.
Hellenic Organization of Agricultural Insurances (ELGA)
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Search repository
Sano, Teruo
Naoi, Takashi
Kashiwagi, Akiko
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| 抄録 |
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内容記述タイプ |
Abstract |
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内容記述 |
Aptamers are powerful tools for detecting and analyzing biomolecules that consist of proteins or nucleic acids. However, their application to aptamers against viroids—highly structured self-replicating RNAs—has not yet been explored. In this study, a magnetic bead- and high-throughput sequencing-based SELEX (MB-HTS-SELEX) protocol for selecting potential aptamers against potato spindle tuber viroid (PSTVd) is presented. Full-length biotinylated-PSTVd RNA was transcribed in vitro, immobilized on streptavidin-coated magnetic beads, and incubated with a library of ~3.32 × 1014 molecules of random single-stranded oligo-DNAs (oligo-ssDNAs) of 20, 30, or 40 nucleotides (L20, L30, or L40, respectively) flanked by primer binding sites for downstream PCR amplification. Simultaneous biotin labeling of the anti-aptamer strand of the resulting double-stranded DNA (dsDNA) amplicons facilitated strand separation using streptavidin-coated magnetic beads. After 10 selection rounds, high-throughput sequencing, followed by bioinformatics analysis of the generated sequences, allowed for the detection of several enriched sequences, representing putative PSTVd-binding aptamers. Subsequent pull-down assays showed that the most abundant oligo-ssDNA in L30 was docked on PSTVd molecules. This combination method may ameliorate the selection of high-affinity aptamers against PSTVd, reduce the number of selection cycles, time, and other costs of aptamer production, thereby promoting future massive and cost-effective viroid detection and characterization. |
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言語 |
en |
| 内容記述 |
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内容記述タイプ |
Other |
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内容記述 |
Keywords: ; aptamer; magnetic beads; high-throughput sequencing; SELEX |
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言語 |
en |
| 書誌情報 |
en : International Journal of Molecular Sciences
巻 27,
号 4,
p. 1831,
発行日 2026
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| 関連情報 |
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識別子タイプ |
DOI |
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関連識別子 |
https://doi.org/10.3390/ijms27041831 |
| ISSN |
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収録物識別子タイプ |
PISSN |
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収録物識別子 |
1661-6596 |
| ISSN |
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収録物識別子タイプ |
EISSN |
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収録物識別子 |
1422-0067 |
| 権利情報 |
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権利情報 |
© 2026 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. |
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言語 |
en |
| 権利情報 |
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権利情報 |
https://creativecommons.org/licenses/by/4.0/ |
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言語 |
en |
| 出版タイプ |
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出版タイプ |
VoR |
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出版タイプResource |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| 出版者 |
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出版者 |
MDPI |
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言語 |
en |
Int. J. Mol. Sci_2026_27(4)_1831.pdf (1.2 MB)
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