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学者姓名：郭绍彬
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Abstract ：
Background: Sexually transmitted infections (STIs) rank among the most prevalent acute infectious conditions and remain a major global public health concern. Notable STI pathogens include Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), and Neisseria gonorrhoeae (NG). Early detection and diagnosis are crucial for controlling the spread of STIs. Results: In this study, we utilized toehold switches integrated with a cell-free system to develop a simple, colorimetric, sensitive, specific and rapid method for the parallel detection of CT, UU, and NG. Target DNA and sensor DNA were transcribed into target trigger RNA and toehold switch sensor RNA respectively, within a cell- free transcription system. The binding of target RNA to the toehold switch RNA activated the switch, subsequently initiating the translation of the downstream lacZ gene. The expressed LacZ protein hydrolyzed the substrate chlorophenol red-beta-D-galactopyranoside (CPRG), resulting in a color change from yellow to purple, which provided a visible colorimetric output. The three screened sensors exhibited excellent orthogonality without any observed cross-reactivity. By enhancing sensitivity with recombinase polymerase amplification (RPA), we reliably detected NG in clinical samples using this method, with no interference from other pathogens. Moreover, we selected high-performance toehold switch sensor for paper-based detection, further enhancing portability. Significance: In summary, this technique enables the simple snd sensitive parallel detection of CT, UU, and NG, generating visible colorimetric results without the need for specialized personnel or sophisticated equipment. Given these advantages, this method holds significant potential as a simple and portable diagnostic tool in resource-limited settings or point-of-care testing (POCT) scenarios.
Keyword ：
Cell-free system Cell-free system Colorimetric Colorimetric Paper-based Paper-based Recombinase polymerase amplification Recombinase polymerase amplification Sexually transmitted infection Sexually transmitted infection Toehold switch Toehold switch
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| GB/T 7714 | Fang, Fengling , Guo, Hongyan , Guo, Zhaopei et al. A simple and colorimetric method utilizing cell-free toehold switch sensors for the detection of Chlamydia trachomatis, Ureaplasma urealyticum and Neisseria gonorrhoeae [J]. | ANALYTICA CHIMICA ACTA , 2025 , 1339 . |
| MLA | Fang, Fengling et al. "A simple and colorimetric method utilizing cell-free toehold switch sensors for the detection of Chlamydia trachomatis, Ureaplasma urealyticum and Neisseria gonorrhoeae" . | ANALYTICA CHIMICA ACTA 1339 (2025) . |
| APA | Fang, Fengling , Guo, Hongyan , Guo, Zhaopei , Lin, Lin , Lai, Lu , Shi, Yue et al. A simple and colorimetric method utilizing cell-free toehold switch sensors for the detection of Chlamydia trachomatis, Ureaplasma urealyticum and Neisseria gonorrhoeae . | ANALYTICA CHIMICA ACTA , 2025 , 1339 . |
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BackgroundDe-novo-designed synthetic transcriptional regulators have great potential as the genetic parts for constructing complex multilayered gene circuits. The design flexibility afforded by advanced nucleic acid sequence design tools vastly expands the repertoire of regulatory elements for circuit design. In principle, the design space of synthetic regulators should allow for the construction of regulatory circuits of arbitrary complexity; still, the orthogonality and robustness of such components have not been fully elucidated, thereby limiting the depth and width of synthetic circuits.ResultsIn this work, we systematically explored the design strategy of synthetic transcriptional regulators, termed switchable transcription terminators. Specifically, by redesigning key sequence domains, we created a high-performance switchable transcription terminator with a maximum fold change of 283.11 upon activation by its cognate input RNA. Further, an automated design algorithm was developed for these elements to improve orthogonality for a complex multi-layered circuit construction. The resulting orthogonal switchable transcription terminators could be used to construct a three-layer cascade circuit and a two-input three-layer OR gate.ConclusionsWe demonstrated a practical strategy for designing standardized regulatory elements and assembling modular gene circuits, ultimately laying the foundation for the streamlined construction of complex synthetic gene circuits.
Keyword ：
Computational design Computational design Multilayered cascades Multilayered cascades Synthetic logic circuits Synthetic logic circuits Synthetic riboregulator Synthetic riboregulator Transcriptional regulation Transcriptional regulation
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| GB/T 7714 | Zhao, Mingming , Kim, Jeongwon , Jiao, Jiayan et al. Construction of multilayered gene circuits using de-novo-designed synthetic transcriptional regulators in cell-free systems [J]. | JOURNAL OF BIOLOGICAL ENGINEERING , 2024 , 18 (1) . |
| MLA | Zhao, Mingming et al. "Construction of multilayered gene circuits using de-novo-designed synthetic transcriptional regulators in cell-free systems" . | JOURNAL OF BIOLOGICAL ENGINEERING 18 . 1 (2024) . |
| APA | Zhao, Mingming , Kim, Jeongwon , Jiao, Jiayan , Lim, Yelin , Shi, Xianai , Guo, Shaobin et al. Construction of multilayered gene circuits using de-novo-designed synthetic transcriptional regulators in cell-free systems . | JOURNAL OF BIOLOGICAL ENGINEERING , 2024 , 18 (1) . |
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The complexity of genetic circuits has not seen a significant increase over the last decades, even with the rapid development of synthetic biology tools. One of the bottlenecks is the limited number of orthogonal transcription factor-operator pairs. Researchers have tried to use aptamer-ligand pairs as genetic parts to regulate transcription. However, most aptamers selected using traditional methods cannot be directly applied in gene circuits for transcriptional regulation. To that end, we report a new method called CIVT-SELEX to select DNA aptamers that can not only bind to macromolecule ligands but also undergo significant conformational changes, thus affecting transcription. The single-stranded DNA library with affinity to our example ligand human thrombin protein is first selected and enriched. Then, these ssDNAs are inserted into a genetic circuit and tested in the in vitro transcription screening to obtain the ones with significant inhibitory effects on downstream gene transcription when thrombins are present. These aptamer-thrombin pairs can inhibit the transcription of downstream genes, demonstrating the feasibility and robustness of their use as genetic parts in both linear DNAs and plasmids. We believe that this method can be applied to select aptamers of any target ligands and vastly expand the genetic part library for transcriptional regulation.
Keyword ：
aptamer aptamer CIVT-SELEX CIVT-SELEX genetic part genetic part synthetic biology synthetic biology thrombin thrombin transcriptional regulation transcriptional regulation
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| GB/T 7714 | Guo, Shaobin , Xu, Zeqi , Lin, Lujie et al. Using CIVT-SELEX to Select Aptamers as Genetic Parts to Regulate Gene Circuits in a Cell-Free System [J]. | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2023 , 24 (3) . |
| MLA | Guo, Shaobin et al. "Using CIVT-SELEX to Select Aptamers as Genetic Parts to Regulate Gene Circuits in a Cell-Free System" . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 24 . 3 (2023) . |
| APA | Guo, Shaobin , Xu, Zeqi , Lin, Lujie , Guo, Yan , Li, Jingying , Lu, Chunhua et al. Using CIVT-SELEX to Select Aptamers as Genetic Parts to Regulate Gene Circuits in a Cell-Free System . | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES , 2023 , 24 (3) . |
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Small molecule aptamers discovered by traditional selection methods usually lack conformational changes upon target binding.This limits the use of aptamers as molecular probes for small molecule detection and regulatory elements of genetic circuits.Here,we report a new method called capture and in vitro transcription-systematic evolution of ligands by exponential enrichment(CIVT-SELEX)to select DNA aptamers that can not only bind to small molecule ligands but also undergo significant con-formational changes.Through this method,we select a structure-switching aptamer of uridine-5'-diphosphate(UDP).Taking advantage of its conformational changes,we first construct a UDP-responsive transcriptional switch by inserting the aptamer in a genetic circuit and demonstrate that it can respond to the addition of UDP and regulate the transcription of downstream genes.We also build a UDP aptamer-based biosensor that can be used for active glycosyltransferase screening.We believe this method can provide a universal platform for selecting small molecule aptamers with conformational changes and expand the use of aptamers in small molecule detection and genetic regulation.
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| GB/T 7714 | Shaobin Guo , Jingjing Lin , Lujie Lin et al. Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors [J]. | 中国科学：化学（英文版） , 2023 , 66 (5) : 1529-1536 . |
| MLA | Shaobin Guo et al. "Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors" . | 中国科学：化学（英文版） 66 . 5 (2023) : 1529-1536 . |
| APA | Shaobin Guo , Jingjing Lin , Lujie Lin , Wen Xu , Yan Guo , Zipeng Xu et al. Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors . | 中国科学：化学（英文版） , 2023 , 66 (5) , 1529-1536 . |
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本发明具体公开一种基于无细胞转录翻译系统的核糖开关，以及在V.vulnificus检测中的应用，实现了无需昂贵仪器设备的V.vulnificus现场检测，且检测结果肉眼可见；本发明中将等温扩增技术RPA与核糖开关相耦合，检测V.vulnificus保守序列的灵敏度为4.30×103copies/μL；经冷冻干燥后的TX‑TL核糖开关检测体系，其活性有所下降，但仍然可实现V.vulnificus保守序列的检测。本发明还通过TX‑TL核糖开关检测体系的重复性试验和特异性试验，证明了该检测方法稳定性好，特异性强，适用于V.vulnificus病原微生物的检测。
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| GB/T 7714 | 郭绍彬 , 金天宇 , 石贤爱 . 一种基于无细胞转录翻译系统的核糖开关与应用 : CN202210647219.5[P]. | 2022-06-09 00:00:00 . |
| MLA | 郭绍彬 et al. "一种基于无细胞转录翻译系统的核糖开关与应用" : CN202210647219.5. | 2022-06-09 00:00:00 . |
| APA | 郭绍彬 , 金天宇 , 石贤爱 . 一种基于无细胞转录翻译系统的核糖开关与应用 : CN202210647219.5. | 2022-06-09 00:00:00 . |
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Abstract ：
Small molecule aptamers discovered by traditional selection methods usually lack conformational changes upon target binding. This limits the use of aptamers as molecular probes for small molecule detection and regulatory elements of genetic circuits. Here, we report a new method called capture and in vitro transcription-systematic evolution of ligands by exponential enrichment (CIVT-SELEX) to select DNA aptamers that can not only bind to small molecule ligands but also undergo significant conformational changes. Through this method, we select a structure-switching aptamer of uridine-5 '-diphosphate (UDP). Taking advantage of its conformational changes, we first construct a UDP-responsive transcriptional switch by inserting the aptamer in a genetic circuit and demonstrate that it can respond to the addition of UDP and regulate the transcription of downstream genes. We also build a UDP aptamer-based biosensor that can be used for active glycosyltransferase screening. We believe this method can provide a universal platform for selecting small molecule aptamers with conformational changes and expand the use of aptamers in small molecule detection and genetic regulation.
Keyword ：
aptamers aptamers biosensors biosensors CIVT-SELEX CIVT-SELEX conformational change conformational change small molecule small molecule
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| GB/T 7714 | Guo, Shaobin , Lin, Jingjing , Lin, Lujie et al. Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors [J]. | SCIENCE CHINA-CHEMISTRY , 2023 , 66 (5) : 1529-1536 . |
| MLA | Guo, Shaobin et al. "Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors" . | SCIENCE CHINA-CHEMISTRY 66 . 5 (2023) : 1529-1536 . |
| APA | Guo, Shaobin , Lin, Jingjing , Lin, Lujie , Xu, Wen , Guo, Yan , Xu, Zipeng et al. Selecting small molecule DNA aptamers with significant conformational changes for constructing transcriptional switches and biosensors . | SCIENCE CHINA-CHEMISTRY , 2023 , 66 (5) , 1529-1536 . |
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Plants have an extensively large number of enzymes including glycosyltransferases that are important in the biosynthesis of natural products. However, it is time-consuming and challenging to study these enzymes and only a small percentage of them have been well-characterized. Here, we report a rapid method to screen plant glycosyltransferases using a linear DNA expression template (LET) based cell-free transcription-translation system (TX-TL). As a proof of concept, we amplified and tested glycosyltransferases from Arabidopsis thaliana and showed that the catalytic activity results of these glycosyltransferases from LET-based-TX-TL were consistent with previous studies. We then chose a local medicinal plant Anoectochilus roxburghii, acquired its transcriptome sequences, and applied this method to study its glycosyltransferases. We rapidly expressed all the putative UDPglucose glycosyltransferases using LET-based-TX-TL and discovered 6 unreported active glycosyltransferases which can catalyze the glycosylation of quercetin into isoquercitrin. Thus, LET-based-TX-TL was shown to be a powerful tool for researchers to rapidly screen plant glycosyltransferases for the first time.
Keyword ：
Anoectochilus roxburghii Anoectochilus roxburghii Cell-free transcription-translation system (TX-TL) Cell-free transcription-translation system (TX-TL) Isoquercitrin Isoquercitrin Linear DNA expression Template (LET) Linear DNA expression Template (LET) Orchidaceae Orchidaceae UDP-Glycosyltransferases UDP-Glycosyltransferases
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| GB/T 7714 | Guo, Shaobin , Wang, Mingdi , Xu, Wen et al. Rapid screening of glycosyltransferases in plants using a linear DNA expression template based cell-free transcription-translation system [J]. | PHYTOCHEMISTRY , 2022 , 193 . |
| MLA | Guo, Shaobin et al. "Rapid screening of glycosyltransferases in plants using a linear DNA expression template based cell-free transcription-translation system" . | PHYTOCHEMISTRY 193 (2022) . |
| APA | Guo, Shaobin , Wang, Mingdi , Xu, Wen , Zou, Fuxian , Lin, Jingjing , Peng, Qin et al. Rapid screening of glycosyltransferases in plants using a linear DNA expression template based cell-free transcription-translation system . | PHYTOCHEMISTRY , 2022 , 193 . |
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Objective To increase the production of (R)-alpha-lipoic acid directly from octanoic acid using engineered Escherichia coli with the regeneration of S-adenosylmethionine.Results The biosynthesis of (R)-alpha-lipoic acid (LA) in E. coli BL21(DE3) is improved by co-expression of lipoate-protein ligase A (LplA) from E. coli MG1655 and lipoate synthase (LipA) from Vibrio vulnificus. The engineered strain produces 20.99 mu g l(-1) of LA in shake flask cultures. The titers of LA are increased to 169.28 mu g l(-1) after the optimization of the medium components and fermentation conditions. We find that the [4Fe-4S] cluster is important for the activity of LipA and co-expression of iscSUA promotes the regeneration of the [4Fe-4S] cluster and leads to the highest LA titer of 589.30 mu g l(-1).Conclusion The method described here can be widely applied for the biosynthesis of (R)-alpha-lipoic acid and other metabolites.
Keyword ：
alpha-Lipoic acid alpha-Lipoic acid Escherichia coli Escherichia coli Metabolic engineering Metabolic engineering Synthesis Synthesis
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| GB/T 7714 | Xiao, Jianbin , Guo, Shaobin , Shi, Xian'ai . Metabolic engineering of Escherichia coli for the production of (R)-a-lipoic acid [J]. | BIOTECHNOLOGY LETTERS , 2022 , 45 (2) : 273-286 . |
| MLA | Xiao, Jianbin et al. "Metabolic engineering of Escherichia coli for the production of (R)-a-lipoic acid" . | BIOTECHNOLOGY LETTERS 45 . 2 (2022) : 273-286 . |
| APA | Xiao, Jianbin , Guo, Shaobin , Shi, Xian'ai . Metabolic engineering of Escherichia coli for the production of (R)-a-lipoic acid . | BIOTECHNOLOGY LETTERS , 2022 , 45 (2) , 273-286 . |
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Gentamicin C1a is an important precursor to the synthesis of etimicin, a potent antibiotic. Wild type Micromonospora purpurea Gb1008 produces gentamicin C1a, besides four other gentamicin C components: C1, C2, C2a, and C2b. While the previously reported engineered strain M. purpurea GK1101 can produce relatively high titers of C1a by blocking the genK pathway, a small amount of undesirable C2b is still being synthesized in cells. Gene genL (orf6255) is reported to be responsible for converting C1a to C2b and C2 to C1 in Micromonospora echinospora ATCC15835. In this work, we identify the genL that is also responsible for the same methylation in Micromonospora purpurea. Based on M. purpurea GK1101, we construct a new strain with genL inactivated and show that no C2b is produced in this strain. Therefore, we successfully engineer a strain of M. purpurea that solely produces gentamicin C1a. This strain can potentially be used in the industrial production of C1a for the synthesis of etimicin.
Keyword ：
gentamicin C1a gentamicin C1a metabolic engineering metabolic engineering Micromonospora purpurea Micromonospora purpurea N-methyltransferase N-methyltransferase
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| GB/T 7714 | Wei, Zeng , Shi, Xianai , Lian, Rong et al. Exclusive Production of Gentamicin C1a from Micromonospora purpurea by Metabolic Engineering [J]. | ANTIBIOTICS-BASEL , 2019 , 8 (4) . |
| MLA | Wei, Zeng et al. "Exclusive Production of Gentamicin C1a from Micromonospora purpurea by Metabolic Engineering" . | ANTIBIOTICS-BASEL 8 . 4 (2019) . |
| APA | Wei, Zeng , Shi, Xianai , Lian, Rong , Wang, Weibin , Hong, Wenrong , Guo, Shaobin . Exclusive Production of Gentamicin C1a from Micromonospora purpurea by Metabolic Engineering . | ANTIBIOTICS-BASEL , 2019 , 8 (4) . |
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