دورية أكاديمية
Cell-Free Biosensors Based on Modular Eukaryotic Riboswitches That Function in One Pot at Ambient Temperature.
| العنوان: | Cell-Free Biosensors Based on Modular Eukaryotic Riboswitches That Function in One Pot at Ambient Temperature. |
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| المؤلفون: | Ogawa A; Proteo-Science Center, Ehime University, 2-5 Bunkyo, Matsuyama ,Ehime 790-8577, Japan., Fujikawa M; Proteo-Science Center, Ehime University, 2-5 Bunkyo, Matsuyama ,Ehime 790-8577, Japan., Onishi K; Proteo-Science Center, Ehime University, 2-5 Bunkyo, Matsuyama ,Ehime 790-8577, Japan., Takahashi H; Proteo-Science Center, Ehime University, 2-5 Bunkyo, Matsuyama ,Ehime 790-8577, Japan. |
| المصدر: | ACS synthetic biology [ACS Synth Biol] 2024 Jul 19; Vol. 13 (7), pp. 2238-2245. Date of Electronic Publication: 2024 Jun 24. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101575075 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2161-5063 (Electronic) Linking ISSN: 21615063 NLM ISO Abbreviation: ACS Synth Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, D.C. : American Chemical Society, c2012- |
| مواضيع طبية MeSH: | Riboswitch*/genetics , Biosensing Techniques*/methods , Cell-Free System* , RNA, Catalytic*/genetics , RNA, Catalytic*/metabolism , Aptamers, Nucleotide*/metabolism , Aptamers, Nucleotide*/genetics , Temperature*, RNA, Messenger/genetics ; RNA, Messenger/metabolism |
| مستخلص: | Artificial riboswitches responsive to user-defined analytes can be constructed by successfully inserting in vitro selected aptamers, which bind to the analytes, into untranslated regions of mRNA. Among them, eukaryotic riboswitches are more promising as biosensors than bacterial ones because they function well at ambient temperature. In addition, cell-free expression systems allow the broader use of these riboswitches as cell-free biosensors in an environmentally friendly manner without cellular limitations. The current best cell-free eukaryotic riboswitch regulates eukaryotic canonical translation initiation through self-cleavage mediated by an implanted analyte-responsive ribozyme (i.e., an aptazyme, an aptamer-ribozyme fusion). However, it has critical flaws as a sensor: due to the less-active ribozyme used, self-cleavage and translation reactions must be conducted separately and sequentially, and a different aptazyme has to be selected to change the analyte specificity, even if an aptamer for the next analyte is available. We here stepwise engineered novel types of cell-free eukaryotic riboswitches that harness highly active self-cleavage and thus require no reaction partitioning. Despite the single-step and one-pot reaction, these riboswitches showed higher analyte dose dependency and sensitivities than the current best cell-free eukaryotic riboswitch requiring multistep reactions. In addition, the analyte specificity can be changed in an extremely facile way, simply by aptamer substitution (and the subsequent simple fine-tuning for giant aptamers). Given that cell-free systems can be lyophilized for storage and transport, the present one-pot and thus easy-to-handle cell-free biosensors utilizing eukaryotic riboswitches are expected to be widely used for on-the-spot sensing of analytes at ambient temperature. |
| فهرسة مساهمة: | Keywords: aptamer; aptazyme; biosensor; cell-free; riboswitch; ribozyme |
| المشرفين على المادة: | 0 (Riboswitch) 0 (RNA, Catalytic) 0 (Aptamers, Nucleotide) 0 (RNA, Messenger) |
| تواريخ الأحداث: | Date Created: 20240624 Date Completed: 20240719 Latest Revision: 20240719 |
| رمز التحديث: | 20240719 |
| DOI: | 10.1021/acssynbio.4c00341 |
| PMID: | 38913391 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 2161-5063 |
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| DOI: | 10.1021/acssynbio.4c00341 |