DRUG DELIVERY
·
2017
·
IF 6.402
Chen-Lin Feng,Yan-Xing Han,Hui-Hui Guo,Xiao-Lei Ma,Zhi-Qiang Wang,Lu-Lu Wang,Wen-Sheng Zheng,Jian-Dong Jiang
Our previous work proved that sequence specific double strand RNA (dsRNA-p21) effectively activated p21 gene expression of colorectal cancer (CRC) cells and consequently suppressed CRC growth. However, efficient delivery system is a significant challenge to achieve sufficient therapy. In this study, a self-assembled HA/PEI/dsRNA-p21 ternary complex (TC-dsRNA-p21) was developed for the tumor-target delivery of dsRNA-p21 into CRC cells. Hyaluronic acid (HA) was introduced to shield the PEI/dsRNA-p21 binary complexes (BC-dsRNA-p21) for reducing the cytotoxicity of PEI and for increasing the tumor-targeted intracellular uptake by cancer cells through HA-CD44 mediated endocytosis. Comparing to the BC-dsRNA-p21, the TC-dsRNA-p21 showed increase in size, decrease in zeta potential, low cytotoxicity as well as high stability in physiological conditions due to the anionic shielding. Confocal microscopy analysis and flow cytometry confirmed that TC-dsRNA-p21 had high transfection efficiency in the CD44-abundant Lovo cells, as compared with binary complex. In vitro physiological experiment showed that, comparing to the control group, the TC-dsRNA-p21 effectively activated the expression of p21 mRNA and P21 protein, causing blockage of cell cycle at G0/G1 phase and suppression of cancer cell proliferation as well as colony formation. Furthermore, in vivo distribution experiment demonstrated that the TC-dsRNA-p21 could effectively accumulate at rectal wall for up to 10 h, following in situ application. These findings indicated that TC-dsRNA-p21 might hold great potential for delivering dsRNA-p21 to treat CRC.
研究领域:
生物医学工程
·
肿瘤学
·
分子生物学
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Haixiang Lv,Xuchao Li,Jiaqing Li,Chang Yu,Qinghong Zeng,Guogui Ning,Hu Wan,Jianhong Li,Kangsheng Ma,Shun He
The cotton aphid Aphis gossypii Glover (Homoptera: Aphididae) is a notorious agricultural pest that has evolved high levels of resistance to various insecticides. Innovative RNA insecticides, as potential green plant protection products, are anticipated to alleviate the significant issue of pest resistance. Herein, a rough-surface hollow mesoporous silica (RHMS)-based gene/insecticide delivery system was constructed for the co-delivery of CYP6CY13 double-stranded RNA (dsRNA) and the chemical insecticide imidacloprid for the effective control of A. gossypii . The hollow mesoporous structure of RHMS facilitates insecticide loading and further combines with dsRNA through electrostatic interactions to form nanoscale complexes, while protecting the dsRNA from complete degradation by nucleases. The screened ds CYP6CY13 fragment was successfully expressed in the L4440-HT1115 (DE3) RNaseIII - system and inhibiting CYP6CY13 expression by RNA interference (RNAi) significantly increased the susceptibility of A. gossypii adults to imidacloprid. The toxicity of the RHMS/IMI/ds CYP6CY13 complex was significantly higher (1.95-fold) than that of imidacloprid. The results of the pot test showed that the effectiveness of the RHMS/IMI/ds CYP6CY13 complex against A. gossypii improved by 19.95 % in 5 days. Therefore, this nanoparticle-mediated gene/insecticide co-delivery system is expected to be an effective tool for pest control.
研究领域:
农业昆虫学
·
分子生物学
·
植物保护
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DOI
高影响因子
JOURNAL OF INSECT PHYSIOLOGY
·
2025
·
IF 2.3
Xu Wu,Wen-Mei Li,Qiang Xu,Xiao-Ping Yi,Da-He Zhang,Ming-Zhe He,Yu-Hong Ren
RNA interference (RNAi) has shown excellent potential for pest management worldwide. However, the application of RNAi against insects can be inefficient and unreliable. Finding a better delivery system is a crucial factor for enhancing RNAi efficacy. Here, we present a novel and effective approach of conjugating double-stranded RNA (dsRNA) with a cell-penetrating disulfide polymer (CPD) to improve dsRNA stability and RNAi efficiency. The fall armyworm (FAW), Spodoptera frugiperda , is a globally agricultural insect. Chitin synthase B ( CHSB ) and methoprene-tolerant ( Met ) genes, which are essential for the development and growth of FAW, were selected as target genes. The CPD was synthesized using a two-step method for dsRNA delivery. The synthesized CPD/dsRNA complex protected dsRNA from nuclease degradation. The biological application of CPD in Sf9 cells demonstrated low cytotoxicity and high cell viability. Moreover, the CPD-loaded dsRNA entered the cells within 6 h. Bioassays of FAW showed that the relative expression levels of the CHSB and Met genes were reduced. Specifically, at 72 h, the relative expression levels of the CHSB and Met genes were 48.14 % and 37.60 % of those in the control group, respectively. The weight and body length of the larvae decreased significantly, and the mortality rate of CPD/ds CHSB reached 30 %. This CPD was demonstrated for the first time to have excellent delivery performance in insects and is expected to become a new and effective tool for pest control, representing a significant advancement in pest management.
研究领域:
昆虫学
·
分子生物学
·
害虫治理
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生物技术
·
RNA干扰技术
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Caiying Zhang,Yuelong Li,Abdus Samad,Hongliang He,Huan Ma,Yang Chen,Tengchuan Jin
Kyasanur Forest disease virus (KFDV), a tick-borne flavivirus prevalent in India, presents a serious threat to human health. KFDV NS3 helicase (NS3hel) is considered a potential drug target due to its involvement in the viral replication complex. Here, we resolved the crystal structures of KFDV NS3hel apo and its complex with three phosphate molecules, which indicates a conformational switch during ATP hydrolysis . Our data revealed that KFDV NS3hel has a higher binding affinity for dsRNA , and its intrinsic ATPase activity was enhanced by dsRNA while being inhibited by DNA . Through mutagenesis analysis, several residues within motifs I, Ia, III, V, and VI were identified to be crucial for NS3hel ATPase activity. Notably, the M419A mutation drastically reduced NS3hel ATPase activity. We propose that the methionine-aromatic interaction between residues M419 and W294, located on the surface of the RNA-binding channel, could be a target for the design of efficient inhibitor probes. Moreover, epigallocatechin gallate (EGCG), a tea-derived polyphenol, strongly inhibited NS3hel ATPase activity with an IC 50 value of 0.8 μM. Our computational docking data show that EGCG binds at the predicted druggable hotspots of NS3hel. Overall, these findings contribute to the development and design of more effective and specific inhibitors.
研究领域:
病毒学
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结构生物学
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药物设计
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Wenjie Wang,Muhammad Adeel Ghafar,Lu Liuyang,Inzamam Ul Haq,Li Cui,Huizhu Yuan,Liande Wang
RNA pesticides have emerged as a promising alternative to conventional chemical pesticides due to their high specificity and minimal environmental impact. However, the instability of RNA molecules in the environment and the challenges associated with their effective delivery to target pests limit their broader application. This study addresses these challenges by developing a dual delivery system using chitosan (CS) and Metal–Organic Frameworks (MOFs) to enhance the delivery and efficacy of double-stranded RNA (dsRNA) and cycloxaprid against Diaphorina citri, a vector of citrus greening disease. The CS-MOF nanoparticles were synthesized and characterized using scanning electron microscopy (SEM) and dynamic light scattering (DLS). Insect bioassays demonstrated that the codelivery system significantly improved insecticidal activity, achieving over 80% mortality in D. citri within 2 days. The results indicate that the encapsulation of dsRNA within MOFs enhances its stability, while the controlled release properties of the nanoparticles improve the efficacy of cycloxaprid. This novel approach shows great potential in overcoming the limitations of RNA pesticides and offers a sustainable solution for pest management in agriculture. Future research should optimize the delivery system, conduct field trials, and explore its applicability to other agricultural pests.
研究领域:
农业科学
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分子生物学
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农药科学
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DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2024
·
IF 4.3
He Wei,Zhang Xinya,Zou Yangxiaoyu,Li Ji,Chang Le,He Yu-Cai,Jin Qiuheng,Ye Jianren
Introduction: Circular RNAs (circRNAs) are endogenous noncoding RNAs (ncRNAs) with transcriptional lengths ranging from hundreds to thousands. circRNAs have attracted attention owing to their stable structure and ability to treat complicated diseases. Our objective was to create a one-step reaction for circRNA synthesis using wild-type T7 RNA polymerase as the catalyst. However, T7 RNA polymerase is thermally unstable, and we streamlined circRNA synthesis via consensus and folding free energy calculations for hotspot selection. Because of the thermal instability, the permuted intron and exon (PIE) method for circRNA synthesis is conducted via tandem catalysis with a transcription reaction at a low temperature and linear RNA precursor cyclization at a high temperature.Methods: To streamline the process, a multisite mutant T7 RNA polymerase (S430P, N433T, S633P, F849I, F880Y, and G788A) with significantly improved thermostability was constructed, and G788A was used.Results: The resulting mutant exhibited stable activity at 45°C for over an hour, enabling the implementation of a one-pot transcription and cyclization reaction. The simplified circRNA production process demonstrated an efficiency comparable to that of the conventional two-step reaction, with a cyclization rate exceeding 95% and reduced production of immunostimulatory dsRNA byproducts.
研究领域:
分子生物学
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生物化学
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合成生物学
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DOI
PubMed
高影响因子
Materials Today Bio
·
2025
·
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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DOI
PubMed
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 19
·4713
·
IF 4.2
Wei He,Qi Geng,Guiying Ji,Ji Li,Dan Wang,Yucai He,Qiuheng Jin,Jianren Ye
The remarkable efficacy of COVID-19 vaccines has established mRNA as a highly promising biomedical technology. However, the adequate application of mRNA therapeutics necessitates additional measures to mitigate the inherent immunogenicity, which is predominantly caused by dsRNA. As a byproduct of the in vitro transcription of mRNA, dsRNA was reported to be originated through several distinct mechanisms, including the extension of 3′ loop-back hairpins, the extension of hybridized abortive transcripts, and promoter-independent transcription. The intricate mechanisms involved pose a dilemma as the reduction in dsRNA results in a concomitant decrease in other critical quality attributes of mRNA. Here, we demonstrate that the promoter binding motifs of T7 RNA polymerase directly impact the production of promoter-independent transcription-based dsRNA. Specifically, the G753A mutation significantly reduces the formation of dsRNA byproducts, which can further combine with modified nucleotides to enhance the effectiveness of dsRNA mitigation and with previously reported high-integrity mutation K389A to minimize side effects. Accordingly, the present study reports a cost-effective approach to synthesize high-purity, less immunostimulatory mRNA by using an engineered T7 RNA polymerase mutant.
研究领域:
分子生物学
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生物技术
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药学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 19
·10286
·
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
·
分子生物学
·
农业科学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Shuai Li,Jianqin Zhang,Hao Wang,Xingyun Zhao,Lin Yang,Lin Chai,Yangyang Liu,Zhangwu Zhao,Bernard Moussian,Abeer El Wakil,Kun Yan Zhu,Zhanghua Zeng,Jianzhen Zhang
RNA interference (RNAi)-based biopesticides, recognized by Science as one of the top ten scientific breakthroughs of 2024, are poised to drive the third revolution in pesticide development. However, the practical applications of RNAi face critical challenges, particularly in overcoming delivery barriers, such as double-stranded RNA (dsRNA) degradation and poor cellular uptake. In this study, we developed a biologically derived melanin nanoparticles (MNP) functionalized with poly- l -lysine (PLL) to deliver dsRNA in Locusta migratoria , a major agricultural pest. The functionalized melanin nanoparticles (MNP-PLL) self-assemble with dsRNA to form MNP-PLL/dsRNA through electrostatic interactions. We evaluated their stability under different environmental conditions, such as in midgut fluid, under various temperature and UV irradiation, demonstrating significantly enhanced resistance to degradation. Bioassays showed substantial improvements in RNAi efficiency and insect mortality. Using fluorescent labeling and pharmacological inhibitors, we further investigated the cellular uptake mechanisms. In contrast to naked dsRNA, chitosan/dsRNA, carbon quantum dots/dsRNA and Spc/dsRNA, MNP-PLL/dsRNA activated three distinct endocytic pathways: clathrin-mediated endocytosis, caveolin-mediated endocytosis, and macropinocytosis, facilitating efficient cellular internalization. Acute toxicity assays confirmed excellent biocompatibility and safety of MNP-PLL to the Western honey bee and zebrafish. This study represents the first application of PLL-functionalized melanin as an efficient nanocarrier to generate MNP-PLL/dsRNA for agricultural pest control, highlighting their strong potential as a next-generation dsRNA delivery platform for sustainable pest management.
研究领域:
农业生物技术
·
纳米生物技术
·
分子昆虫学
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RNA干扰
·
害虫管理
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产品关联
DOI
高影响因子
MOLECULAR THERAPY
·
2025
·
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
·
分子生物学
·
疫苗开发
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产品关联
DOI
PubMed
高影响因子
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY
·
2019
·
IF 2.142
Rong Xiong,Wei Zhao,Xi Chen,Tian Li,Haiyin Li,Yan Li,Wangshang Shen,Ping Chen
Serotonin is involved in the regulation of many physiological and behavioral processes in vertebrates and invertebrates. The effects of serotonin are mediated through interactions of several 5-HT receptor types. The expression and pharmacological properties of 5-HT1 have received more attention than other serotonin receptors, but its functions at the individual level are little studied in arthropods. Silkworm , a Lepidoptera model, almost has no reports about serotonin receptors. To analyze the function of Bm5-HT1A receptor in vitro , the ORF of Bm5-HT1A was cloned into the pcDNA3.1 vector and expressed in HEK 293 cells . Serotonin activation of Bm5-HT1A -expressing cells decreased forskolin-stimulated cAMP synthesis and had the most potent effect compared to other biogenic amines. Serotonin reduced cAMP synthesis in a dose-dependent manner, and half-maximal activation (EC50) occurred at a concentration of 1.17 × 10 −7 M (117 nM). The pharmacological analysis demonstrated that the rank potency of agonists was pimozide >8-OH-DPAT >5-MeOT ~ αm-5-HT, and antagonists was WAY-100635 > prazosin > SB-269970 > methiothepin at the Bm5-HT1A receptor. Injecting the antagonist of Bm5-HT1A receptor into larvae caused slow or weak motility, and adults lowered courtship vitality or moving speed. Injecting dsRNA of Bm5-HT1A into adults also dropped locomotivity in courtship. These results show that the Bm5-HT1A receptor is related to locomotor activity. This study provides the first information of serotonin receptor on pharmacological in silkworm and on individual functions in arthropods.
研究领域:
神经生物学
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药理学
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昆虫生理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）