Arabian Journal of Chemistry
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2022
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IF 6.212
Jie Zhao,Yan Gong
RNA plays a crucial role in biochemical processes. RNAs with abasic sites (AP-RNA), as one of RNA lesions, will cause serious negative consequences. Efficient and sensitive detection of AP-RNA deserves but does not receive enough attention. Water-soluble uridine triphosphate (UTP)-capped Mn-doped ZnS quantum dots (QDs), which have excellent room-temperature phosphorescence (RTP) properties, were prepared by wet-chemical method. The UTP on the surface of QDs could form a precipitation complex with amiloride (AMI) to quench the RTP. Meanwhile, abasic double-stranded RNA (AP-dsRNA) with a high affinity to AMI could dissociate AMI from the surface of UTP-QDs, resulting in RTP recovery. In this process, a highly sensitive and selective RTP-detection system for AP-dsRNA was constructed with AMI as the modulation factor and UTP-QDs as the luminescence matrix. RTP was turned off and on for the determination of AP-dsRNA, and the interference of background fluorescence and scattered light was effectively avoided during complex formation and aggregation. Steric hindrance of the conformation of AP-dsRNA provided efficient selectivity. The detection limit for AP-dsRNA was 0.86 nM, the relative standard deviation was 2.1%, and the recovery of biological samples with AP-dsRNA addition ranged from 95% to 103% at optimal conditions. The system is expected to provide new methods for environmental monitoring, early disease screening, and evaluation of gene editing efficiency.
研究领域:
生物化学
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分子生物学
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分析化学
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产品关联
DOI
高影响因子
MOLECULAR THERAPY
·
2025
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IF 12
Jingjing Zhang,Chao Li,Yuheng Liu,Rui Liao,Dian He,Lifeng Xu,Tingting Chen,Qin Xiao,Mingxing Luo,Yang Chen,Yali Li,Huaxing Zhu,Joseph Rosenecker,Xiaoyan Ding,Shuchen Pei,Shan Guan
The development of mucosal mRNA vaccines is promising but extremely challenging. Major efforts have been focused on optimizing delivery systems, whereas it is still unknown whether the intrinsic quality of mRNA components significantly impacts the potency of airway inoculated mRNA vaccines. Here, we systematically demonstrate mucosal mRNA vaccine requires higher standards of purification and tailor-designed sequence to fulfil its potency compared to parenteral-route inoculated counterpart. Double-stranded RNA (dsRNA) contaminants are prone to trigger innate immune response in the airway that activates the mRNA degradation mechanism, thereby diminishing the mRNA expression and subsequent antigen-specific immune responses. To address these challenges, we developed a strategy that combines optimized untranslated regions (UTRs) screened from endogenous genes of pulmonary cells with affinity chromatography-based purification which effectively removed dsRNA contaminants. The optimized mRNA administered via airway route not only demonstrated superior protein expression (30-fold increase) and reduced inflammation in the lung, but also promoted robust immunity comprising significantly elevated systemic, cellular, and mucosal immune responses, which was in stark contrast to intramuscular injected counterpart that displays less pronounced benefits. Our findings offer new insights into the development of mucosal mRNA therapeutics from an overlooked but crucial perspective of optimizing mRNA components.
研究领域:
免疫学
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分子生物学
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疫苗开发
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 19
·10286
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IF 4.9
Sen Fan,Yanguang Zhou,Na Zhu,Qingling Meng,Yujin Zhao,Jingyan Xu,Yunjia Tang,Shijie Dai,Xiaofeng Yuan
Fusarium oxysporumis a widespread soil-borne fungal pathogen that can infect various plants, causing wilt and root rot diseases. The root rot disease ofAtractylodes macrocephalacaused byF. oxysporumis among the most serious diseases associated with continuous cropping, significantly hindering its sustainable development. In this study, we aimed to investigate the effect of exogenous application of double-stranded RNA (dsRNA) on silencing theF. oxysporum Tup1gene to reduce its virulence and to evaluate its potential application in controlling root rot disease inA. macrocephala. TheTup1gene was amplified from theF. oxysporumgenome, and different lengths ofTup1-dsRNA were designed and synthesized. The uptake of dsRNA by the fungus was verified usingTup1-dsRNA labeled with fluorescein, and in vitro dsRNA treatment experiments were conducted to assess its impact on the growth and virulence ofF. oxysporum. Additionally,Tup1-dsRNA was applied to the roots ofA. macrocephalato evaluate its effectiveness in controlling root rot disease. The experimental results showed thatF. oxysporumcould effectively uptake exogenously appliedTup1-dsRNA, significantly reducingTup1gene expression. All lengths ofTup1-dsRNA inhibited fungal growth and caused morphological changes in the fungal hyphae. Further plant experiments and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis indicated thatTup1-dsRNA treatment significantly reduced the incidence of root rot disease inA. macrocephala, which was supported by the reduction in peroxidase (POD) and catalase (CAT) enzyme activities, malondialdehyde (MDA) content, and proline (Pro) levels in treated root tissues. This study demonstrated that exogenous dsRNA could reduce the virulence ofF. oxysporumby silencing theTup1gene and effectively mitigate the root rot disease it causes inA. macrocephala. The successful application ofTup1-dsRNA provided strong evidence for the potential of RNA interference (RNAi) technology in plant disease control. Future research could further optimize the design and application of dsRNA to enhance its practical value in agriculture.
研究领域:
植物病理学
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分子生物学
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农业科学
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产品关联
DOI
PubMed
高影响因子
Jove-Journal of Visualized Experiments
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2022
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IF 1.424
Xiaofang Liu,Xiang Zhou,Lifeng Zhou,Jiafu Hu,Kai Guo
The pinewood nematode, Bursaphelenchus xylophilus, is one of the most destructive invasive species worldwide, causing the wilting and eventual death of pine trees. Despite the recognition of their economic and environmental significance, it has thus far been impossible to study the detailed gene functions of plant-parasitic nematodes (PPNs) using conventional forward genetics and transgenic methods. However, as a reverse genetics technology, RNA interference (RNAi) facilitates the study of the functional genes of nematodes, including B. xylophilus. This paper outlines a new protocol for RNAi of the ppm-1 gene in B. xylophilus, which has been reported to play crucial roles in the development and reproduction of other pathogenic nematodes. For RNAi, the T7 promoter was linked to the 5′-terminal of the target fragment by polymerase chain reaction (PCR), and double-stranded RNA (dsRNA) was synthesized by in vitro transcription. Subsequently, dsRNA delivery was accomplished by soaking the nematodes in a dsRNA solution mixed with synthetic neurostimulants. Synchronized juveniles of B. xylophilus (approximately 20,000 individuals) were washed and soaked in dsRNA (0.8 µg/mL) in the soaking buffer for 24 h in the dark at 25 °C.The same quantity of nematodes was placed in a soaking buffer without dsRNA as a control. Meanwhile, another identical quantity of nematodes was placed in a soaking buffer with green fluorescent protein (gfp) gene dsRNA as a control. After soaking, the expression level of the target transcripts was determined using real-time quantitative PCR. The effects of RNAi were then confirmed using microscopic observation of the phenotypes and a comparison of the body size of the adults among the groups. The current protocol can help advance research to better understand the functions of the genes of B. xylophilus and other parasitic nematodes toward developing control strategies through genetic engineering.
研究领域:
分子生物学
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植物病理学
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线虫学
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遗传学
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产品关联
DOI
PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2023
·
IF 19
Zhiwen Wang,Yu Li,Borui Zhang,Xiang Gao,Mengru Shi,Sicong Zhang,Shan Zhong,Yang Zheng,Xili Liu
Spray-induced gene silencing (SIGS) represents an attractive avenue for plant protection, but limited uptake efficiency of double-stranded RNA (dsRNA) has restricted its application against the notorious oomycetes, Phytophthora . To control Phytophthora , dsRNAs targeting two essential genes in Phytophthora are screened for their ability to partially decrease Phytophthora capsici ( P. capsici ) infection and fecundity. To further facilitate SIGS for Phytophthora control, functionalized carbon dots (CDs) are complexed with the screened dsRNAs (dsRNA-CDs) through electrostatic interaction. dsRNA-CDs significantly enhanced the control efficacy of dsRNA against Phytophthora infestans , Phytophthora sojae , and both the wild-type and fungicide-resistant P. capsici . The synergism is based on enhancing dsRNA stability and internalization. Dual treatment with dsRNA-CDs and the corresponding fungicide reduced the amount of fungicide required to achieve the same protective effect by 90%. Plant RdRP1 is the main effector in processing various lengths of small RNAs to induce translation inhibition in Phytophthora . Notably, here the first application of a nano-delivery system to improve the effect of SIGS against Phytophthora pathogens is reported. Moreover, the elucidation of how CD facilitates dsRNA internalization in recipient cell and the molecular mechanism of SIGS can benefit the application of dsRNA-CDs in other plant-pathogen pathosystems through improving dsRNA bioactivity and reducing chemical fungicides application.
研究领域:
植物病理学
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分子生物学
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纳米技术
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农业科学
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DOI
高影响因子
Materials Today Bio
·
2025
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IF 10.2
Xiaojie Ding,Xinchun Li,Yan Li,Huiyan Guo,Kexin Cao,Mengnan An,Chong Zhang,Yuanhua Wu,He Liu,Rujun Zhou
Rhizoctonia solani Kühn is an important fungal pathogen that causes serious crop yield losses worldwide. The application of nanoscale strategies based on RNA interference (RNAi) represents an environmentally friendly and efficient approach for plant disease control. In this study, R. solani glycosyl hydrolase family 1 ( RsGH1 ), which functions as a cell wall degrading enzyme (CWDE), and was screened as a prospective RNAi target gene for managing R. solani AG3 TB. Additionally, a novel nanosystem for loading and stabilization of double-stranded RNA (dsRNA) was developed. When the mass ratio of ε-poly-L-lysine (ε-PL) to carboxymethyl chitosan (CMCS) is 1:1, ε-PL can spontaneously conjugate with CMCS to form nanoscale spherical particles by electrostatic interaction, hydrogen bonding and Van der Waals forces. Compared with ε-PL alone, the introduction of CMCS resulted in more regular and uniform nanoparticles. ds RsGH1 spontaneously binds with ε-PL@CMCS, which is referred to as ds RsGH1 @ε-PL@CMCS. ε-PL@CMCS protected dsRNA from RNase A degradation effectively. The combination of ds RsGH1 with ε-PL@CMCS remarkably improved the deposition and adhesion of ds RsGH1 droplets on Nicotiana tabacum ( N. tabacum ) leaves. The application of ε-PL@CMCS improved the RNAi efficiency of ds RsGH1 and prolonged its protective duration on crops. ds RsGH1 derived from R. solani AG3 TB also exhibited broad-spectrum activity against R. solani AG1-IA in rice and maize plants. In this study, a self-assembled multi-component nano-fungicide was designed based on dsRNA and nanocarriers. This work proposes an eco-friendly strategy to manage R. solani .
研究领域:
纳米技术
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植物病理学
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真菌学
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RNA生物学
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农业生物技术
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材料科学
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产品关联
DOI
PubMed
高影响因子
POLYHEDRON
·
2025
·
IF 2.6
Keliang Wan,Lifeng Tan
Double-stranded RNA determines and regulates many biological processes in cells, plays a vital role in cell information transmission and gene regulation, and is a key target for the treatment of certain diseases. Here, we synthesized and characterized two novel ruthenium(II) complexes [Ru(phen) 2 ( o -CNPIP)] 2+ (Ru 1 ) and [Ru(phen) 2 ( o -CH 3 PIP)] 2+ (Ru 2 , phen = 1,10-phenanthroline, o -CNPIP = (2-(4-cyanodipyrido)-imidazo-[4,5- f ][1,10]-phenanthroline, o -CH 3 PIP = 2-(4-methyl)-imidazo-[4,5- f ][1,10]-phenanthroline). The interaction of the two Ru (II) complexes with double-stranded RNA poly(rA)•poly(rU) under dilute solution and molecular crowding conditions was studied by various spectroscopic methods and viscosity measurements. The results of UV–vis absorption spectroscopy and steady-state emission spectroscopy studies show that Ru 1 showed stronger binding and stabilizing ability to poly(rA)•poly(rU) and the binding affinity of Ru 1 and Ru 2 to poly(rA)•poly(rU) was weakened under molecular crowding conditions. Subsequently, the results of circular dichroism spectroscopy studies, thermal denaturation measurements and viscosity measurements also confirmed the results of our UV–vis absorption spectroscopy and steady-state emission spectroscopy studies. The results also showed that the two Ru(II) complexes bind, stabilize and perturb the structure of poly(rA)•poly(rU) by intercalation under dilute solution and molecular crowding conditions. In addition, it was found that the strength of these abilities was closely related to the binding affinity of the Ru(II) complexes to poly(rA)•poly(rU) by comparing the results of these studies. Furthermore, the thermodynamic parameters for the binding of Ru(II) complexes to poly(rA)•poly(rU) under the two conditions indicate that the binding process is spontaneous, is an enthalpy-driven process, and that hydrogen bonding serves as the dominant force. The purpose of this work is to explore the substituent effect of Ru(II) complexes and the effect of solution environment on the ability of Ru(II) complexes to bind and stabilize poly(rA)•poly(rU). Some insights resulting from this work are expected to facilitate the design of highly selective double-stranded RNA binders based on Ru(II) complexes. Interestingly, the phosphorescence intensity of Ru 2 after binding to poly(rA)•poly(rU) is significantly higher than that of Ru 1 under both solution conditions, with even stronger phosphorescence emission observed under molecular crowding conditions. This indicates that Ru 2 possesses the potential to serve as a nucleic acid luminescent probe for nucleic acid imaging in biological cells. To enhance its luminescent properties and specific binding capacity, further structural modification of Ru 2 is essential, which will be one of the key focuses of our future research efforts.
研究领域:
无机化学
·
生物无机化学
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核酸化学
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分子光谱学
·
生物物理化学
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DOI
高影响因子
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