MOLECULAR PLANT PATHOLOGY
·
2024
·Vol. 25
·期 1
·e13416
·
IF 4.8
Wanyue Li,Zeming Liu,Yuli Huang,Jie Zheng,Yang Yang,Yimeng Cao,Liwen Ding,Yuling Meng,Weixing Shan
Phytophthora infestans is a destructive oomycete that causes the late blight of potato and tomato worldwide. It secretes numerous small proteins called effectors in order to manipulate host cell components and suppress plant immunity. Identifying the targets of these effectors is crucial for understanding P. infestans pathogenesis and host plant immunity. In this study, we show that the virulence RXLR effector Pi23014 of P. infestans targets the host nucleus and chloroplasts. By using a liquid chromatogrpahy-tandem mass spectrometry assay and co-immunoprecipitation assasys, we show that it interacts with NbRBP3a, a putative glycine-rich RNA-binding protein. We confirmed the co-localization of Pi23014 and NbRBP3a within the nucleus, by using bimolecular fluorescence complementation. Reverse transcription-quantitative PCR assays showed that the expression of NbRBP3a was induced in Nicotiana benthamiana during P. infestans infection and the expression of marker genes for multiple defence pathways were significantly down-regulated in NbRBP3 -silenced plants compared with GFP -silenced plants. Agrobacterium tumefaciens -mediated transient overexpression of NbRBP3a significantly enhanced plant resistance to P. infestans . Mutations in the N-terminus RNA recognition motif (RRM) of NbRBP3a abolished its interaction with Pi23014 and eliminated its capability to enhance plant resistance to leaf colonization by P. infestans . We further showed that silencing NbRBP3 reduced photosystem II activity, reduced host photosynthetic efficiency, attenuated Pi23014-mediated suppression of cell death triggered by P. infestans pathogen-associated molecular pattern elicitor INF1 , and suppressed plant immunity.
研究领域:
植物病理学
·
分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Xiaojiao Zhu,Gang Liu,Yingcui Bu,Jie Zhang,Lianke Wang,Yupeng Tian,Jianhua Yu,Zhichao Wu,Hongping Zhou
Cell viability is greatly affected by external stimulus eliciting correlated dynamical physiological processes for cells to choose survival or death. A few fluorescent probes have been designed to detect whether the cell is in survival state or apoptotic state, but monitoring the regulation process of the cell undergoing survival to death remains a long-standing challenge. Herein, we highlight the in situ monitor of mitochondria regulating the cell viability by the RNA-specific fluorescent photosensitizer L. At normal conditions, L anchored mitochondria and interacted with mito-RNA to light up the mitochondria with red fluorescence. With external light stimulus, L generated reactive oxide species (ROS) and cause damage to mitochondria, which activated mitochondrial autophagy to prevent death, during which the red fluorescence of L witnessed dynamical distribution in accordance with the evolution of vacuole structures containing damaged mitochondria into autophagosomes. However, with ROS continuously increasing, the mitochondrial apoptosis was eventually commenced and L with red fluorescent was gradually accumulated in the nucleoli, indicating the programmed cell death. This work demonstrated how the delicate balance between survival and death are regulated by mitochondria.
研究领域:
细胞生物学
·
分子生物学
·
生物医学工程
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Yi Chen,Hengyi Chen,Xiaohui Chen,Yufang Zhang,Wei Tu,Jiaxi Mo,Shanshan Tian,Chenyu Sun,Xiaoqin Meng,Zhizeng Wang,Jichun Yang,Yang Luo
Accurate intracellular visualization of human telomerase RNA (hTR) is imperative for early diagnosis and treatment monitoring of hepatocellular carcinoma (HCC). While isothermal amplification-based DNA cascade strategies are promising, challenges persist in achieving great intake efficiency of detection probes within tumor cells and enhancing intracellular reaction efficiency. This study introduces a SA@Comb-HCR nanosystem , a highly effective approach for in situ hTR detection in HCC cells. Sodium alginate-coated liposomes ensures efficient nanoprobe delivery, which are then combined with proximity effect-inspired signal amplification . The coating of sodium alginate facilitates receptor-mediated endocytosis , prevents serum protein adhesion, and mitigates cationic liposome cytotoxicity. The designed Comb-like consolidated hairpin probe enhances the concentration of the local reactant, resulting in cascade amplification upon hTR activation. This technique achieves precision detection of intracellularly overexpressed hTR in HCC cells with a remarkable detection limit of 0.7 pM. This approach holds great promise for advancing targeted and sensitive early clinical diagnosis of HCC.
研究领域:
分子生物学
·
肿瘤学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
Journal of the American Chemical Society
·
2025
·
IF 15.6
Yiming Liu,Mingrui Zhang,Jingjing Zhao,Yongshuo Ren,Shubin Li,Weichen Wang,Wei Mu,Xiaojun Han
The reconstitution of the de novo nucleotide synthesis pathway from inorganic compounds for RNA transcription is essential for an autonomous artificial cell. Here, a de novo uridine triphosphate (UTP) synthesis pathway is successfully constructed starting from NH4HCO3, involving eight enzymes of carbamoyl phosphate synthetase (CPS), aspartate transcarbamoylase (ATC), dihydroorotase (DHO), dihydroorotate dehydrogenase (DHODH), ribose-phosphate pyrophosphokinase (RPPK), uridine 5′-monophosphate synthase (UMPs), uridylate kinase (UK), and nucleoside diphosphate kinase (NDK). ATP regeneration system composed of creatine kinase (CK) and creatine phosphate is incorporated to drive this pathway. 0.85 mM UTP is produced within 180 min under the optimized conditions, which is further used for RNA transcription along with preadded CTP, GTP, regenerated ATP, and T7 RNA polymerase. The de novo UTP synthesis pathway and RNA transcription components are encapsulated into giant unilamellar vesicles to build artificial cells capable of nucleotide de novo production for subsequent RNA transcription. The successful RNA transcription inside artificial cells is visualized using pyronin Y. The successful reconstitution of the de novo UTP synthesis pathway for RNA transcription inside artificial cells lays a solid foundation for the reproduction of an autonomous artificial cell.
研究领域:
合成生物学
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生物化学
·
分子生物学
·
人工细胞
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系统生物学
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2024
·Vol. 15
·期 1
·1-15
·
IF 14.7
Zhou Zhixin,Han Bing,Wang Yu,Lin Nina,Zhou Zhongqiu,Zhang Yuan,Bai Ying,Shen Ling,Shen Yanfei,Zhang Yuanjian,Yao Honghong
While circular RNAs (circRNAs) exhibit lower abundance compared to corresponding linear RNAs, they demonstrate potent biological functions. Nevertheless, challenges arise from the low concentration and distinctive structural features of circRNAs, rendering existing methods operationally intricate and less sensitive. Here, we engineer an intelligent tetrahedral DNA framework (TDF) possessing precise spatial pattern-recognition properties with exceptional sensing speed and sensitivity for circRNAs. The signal output of TDF sensor occurs only when multivalent spatial pattern-recognition of a circRNA in unamplified samples. Using this sensor, we visualize the real-time response of endogenous circRNA expression in vitro neuronal cells and in vivo brain between pre-stroke and post-stroke male mice, identify the patients with acute ischemic stroke in clinical samples, as well as track the delivery of circRNA in photochromic stroked animal model. Thus, the TDF sensor provides a fast and sensitive tool for the detection of circRNA abundance in both physiological and pathophysiological conditions.
研究领域:
分子生物学
·
生物医学工程
·
神经科学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2022
·
IF 3.322
Ye Gao,He Xin,Bolei Cai,Le Wang,Qianxin Lv,Yan Hou,Fuwei Liu,Taiqiang Dai,Liang Kong
RNA interference (RNAi)-based gene therapy that promotes anabolic bone formation is an effective approach for addressing osteoporosis. However, the selection of target gene and tissue-specific delivery systems has hindered the progression of this strategy. In this study, we identified casein kinase-2 interacting protein-1 encoding gene ( Ckip-1 ), a negative regulator of bone formation, as an effective target of small interfering RNAs (siRNAs) for improving bone mass. Moreover, an impressive (DSS) 6 -Liposome (Lipos) nanoparticle system that could target the bone formation surface was synthesized to enhance the delivery of Ckip-1 siRNA to osteogenic lineage cells. The in vitro results confirmed that the (DSS) 6 -Lipos system could efficaciously improve the intracellular delivery of Ckip-1 siRNA without obvious cell toxicity. The in vivo application of the delivery system showed specific accumulation of siRNA in osteogenic cells located around the bone formation surface. Bone-related analysis indicated increased bone mass and improved bone microarchitecture in mice with ovariectomy-induced osteoporosis. Moreover, the biomechanical characteristics of the tibia were enhanced significantly, indicating increased resistance to fragile fracture induced by osteoporosis. Thus, (DSS) 6 -Lipos- Ckip-1 siRNA-based osteoanabolic therapy may be a promising option for the treatment of osteoporosis.
研究领域:
药理学
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基因治疗
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骨生物学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Ying Peng,Yanhong Pan,Zhaowei Sun,Jinlong Li,Yongxiang Yi,Jie Yang,Genxi Li
The pandemic of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is continuously worsening globally, herein we have proposed an electrochemical biosensor for the sensitive monitoring of SARS-CoV-2 RNA. The presence of target RNA firstly triggers the catalytic hairpin assembly circuit and then initiates terminal deoxynucleotidyl transferase-mediated DNA polymerization. Consequently, a large number of long single-stranded DNA products can be produced, and these negatively charged DNA products will bind a massive of positively charged electroactive molecular of Ru(NH 3 ) 6 3+ due to the electrostatic adsorption. Therefore, significantly amplified electrochemical signals can be generated for sensitive analysis of SARS-CoV-2 RNA in the range of 0.1–1000 pM with the detection limit as low as 26 fM. Besides the excellent distinguishing ability for SARS-CoV-2 RNA against single-base mismatched RNA, the proposed biosensor can also be successfully applied to complex matrices, as well as clinical patient samples with high stability, which shows great prospects of clinical application.
研究领域:
分子生物学
·
分析化学
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
IMMUNITY
·
2024
·
IF 25.5
Rui Ding,Xiaoyan Yu,Zhilin Hu,Yu Dong,Haiyan Huang,Yuerong Zhang,Qiaoqiao Han,Zhi-Yu Ni,Ren Zhao,Youqiong Ye,Qiang Zou
RNA splicing is involved in cancer initiation and progression, but how it influences host antitumor immunity in the metabolically abnormal tumor microenvironment (TME) remains unclear. Here, we demonstrate that lactate modulates Foxp3-dependent RNA splicing to maintain the phenotypic and functional status of tumor-infiltrating regulatory T (Treg) cells via CTLA-4. RNA splicing in Treg cells was correlated with the Treg cell signatures in the TME. Ubiquitin-specific peptidase 39 (USP39), a component of the RNA splicing machinery, maintained RNA-splicing-mediated CTLA-4 expression to control Treg cell function. Mechanistically, lactate promoted USP39-mediated RNA splicing to facilitate CTLA-4 expression in a Foxp3-dependent manner. Moreover, the efficiency of CTLA-4 RNA splicing was increased in tumor-infiltrating Treg cells from patients with colorectal cancer. These findings highlight the immunological relevance of RNA splicing in Treg cells and provide important insights into the environmental mechanism governing CTLA-4 expression in Treg cells.
研究领域:
免疫学
·
肿瘤学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Radiation Research and Applied Sciences
·
2024
·
IF 1.7
Zheng Feng,Zhouzhan Luo
This work aimed to evaluate the therapeutic efficacy of clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleic acid (RNA) nanoparticles (NPs) in the therapy of cervical cancer (CC) using ultrasound elastography imaging. Using poly(β-amino esters) (PBAE), CRISPR RNA NPs encapsulating Human Papillomavirus 16 (HPV16) E7, namely Poly(β-amino ester) (PBAE)/CRISPR16E7-GFP and PBAE/shRNA-16E7, were prepared. The characterization of these NPs was conducted using transmission electron microscopy (TEM) and laser diffraction particle size analysis. The cytotoxicity of PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs on CC cell lines SiHa, HeLa, and CaSki was assessed using the cell counting kit-8 (CCK-8) assay. Sixty female Sprague Dawley (SD) rats were rolled into six groups randomlyas follows: the blank Ctrl group (Group A, no treatment), the model group (Group B, CC model), the pSpCas9-GFP plasmid group (Group C, CC model + pSpCas9-GFP), the PBAE/CRISPR-16E7 group (Group D, CC model + PBAE/CRISPR-16E7), the pSIREN-U6-shRNA-CMV-iRFP plasmid group (Group E, CC model + pSIREN-U6-shRNA-CMV-iRFP), and the PBAE/shRNA-16E7 group (Group F, CC model + PBAE/shRNA-16E7), each consisting of 10 rats. The impact of NPs on the expression levels (ELs) of HPV16E7 and RB1 proteins in tumor tissues was assessed using Western Blot (WB) analysis. Additionally, ultrasound elastography imaging was employed to analyze the strain ratio (SR) and shear wave velocity (SWV) in lesions of various rat groups. The results indicated that the PBAE/CRISPR16E7-GFP NPs exhibited a particle size of 125.64 ± 7.8 nm, a zeta potential of 19.17 ± 3.61 mV, and a polydispersity index (PDI) of 0.281 ± 0.03. These NPs demonstrated a regular spherical structure with a compact morphology. The PBAE/shRNA-16E7 NPs displayed a particle size of 112.93 ± 9.1 nm, a zeta potential of 13.54 ± 1.39 mV, and a PDI of 0.185 ± 0.03. They exhibited a uniform distribution of spherical shape with regularity in size. The plasmids within PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs exhibited distinct trends of burst and sustained release. The viability of SiHa, HeLa, and CaSki cells remained unaffected by PBAE/CRISPR16E7-GFP NPs. At various mass ratios, neglectable differences ( P > 0.05) were observed in cell viability of SiHa, HeLa, and CaSki cells treated with PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 NPs relative to Ctrl group. Relative to Group B, both Groups C and E exhibited a drastic decrease in tumor volume, tumor mass, and HPV16E7 protein ELs ( P < 0.05), whereas Groups D and F showed a notably greater reduction in tumor volume, tumor mass, and HPV16E7 protein ELs ( P < 0.01). The HPV16E7 protein ELs, SR, and SWV values greatly increased ( P < 0.001) in tumors of Group B rats while the EL of RB1 protein notably decreased ( P < 0.001) versus Group A. Groups C and E demonstrated elevated HPV16E7 ELs, SR, and SWV values ( P < 0.05), with a marked decrease in RB1 protein EL ( P < 0.001). Group F exhibited a notable reduction in HPV16E7 protein levels, SR, and SWV values ( P < 0.001).The CRISPRRNA NPs composed of PBAE encapsulating HPV16 E7, namely PBAE/CRISPR16E7-GFP and PBAE/shRNA-16E7 plasmids, demonstrated a significant role in CC therapy. These NPs hold potential application value in the treatment of HPV-related CC, indicating their importance in addressing this particular disease.
研究领域:
分子生物学
·
肿瘤学
·
医学影像学
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产品关联
DOI
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2022
·Vol. 32
·期 20
·2112023
·
IF 19.924
Pan Cheng,Ke Liu,Yucai Wan,Wei Hu,Cancan Ji,Peng Huang,Qihao Guo,Jia Xu,Qin Cheng,Dong Wang
Engineering 3D nanofibrous monolithic materials with a controllable porous channel structure and chemical activity is promising for the rapid and efficient separation of environmentally sensitive biomacromolecules such as RNA. Herein, freeze-drying method via regulating the viscosity of nanofiber dispersions is developed to fabricate nanofibrous sponges (NFSs) with 3D structures of microsphere, lamella, and honeycomb by combining poly(vinyl alcohol- co -ethylene) (PVA- co -PE) nanofibers with chitosan (CS) and polyethyleneimine (PEI) in dispersions. Among sponges, a honeycomb nanofibrous sponge (HNFS) possesses highly oriented open-cells and optimal performance, including water permeability of 7.431 × 10 3 L h m −2 at 1 psi, static saturated capacity of 887.6 mg g −1 , dynamic capacity of 763.2 mg g −1 at a flow rate of 1 mm min −1 , and mechanical robustness, superior to reported state-of-the-art ion-exchange chromatography materials. Furthermore, HNFS-packed chromatographic, funnel, and centrifugal columns present exceptional performance and broad applicability in rapid-separation of RNA from model solution and diverse organisms without degradation. They are attributed to the highly oriented open-cell structure and high-density adsorption ligands on the surface of nanofibrous cell walls. This work provides a facile approach to design advanced NFSs with versatile physicochemical structure and outstanding performance.
研究领域:
生物工程
·
材料科学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Kaiwu Zhang,Xuan Wang,Caicheng Long,Jin Xu,Zixin Jiang,Bo Feng,Peng Zhang,Junjie Fei,Taiping Qing
DNA templated fluorescent copper nanoclusters (DNA-CuNCs) have demonstrated enormous potential for biochemical analysis. However, most of them are focused on DNA-related sensing. Here, we designed a hairpin DNA/RNA chimeric chain for the formation of fluorescent CuNCs and further applied it to detect reverse transcription-associated ribonuclease H (RNase H). The DNA/RNA chimera templated copper nanoclusters (DNA/RNA-CuNCs) exhibited similar spectral properties with DNA-CuNCs. In the presence of RNase H, the RNA segment in chimera is hydrolyzed and release short oligonucleotides, which fail to synthsize fluorescent CuNCs. The capabilities for RNase H detection from complex samples and screening of inhibitors were verified. This strategy exhibited high sensitivity for RNase H with a detectable minimum concentration of 5.5 × 10 −4 U mL -1 , which is lower than the developed fluorescent methods. This work will open up new thoughts to design new template for metal nanoclusters and broaden their application in RNA-related sensing.
研究领域:
生物化学
·
分子生物学
·
分析化学
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产品关联
DOI
高影响因子
FISH & SHELLFISH IMMUNOLOGY
·
2019
·
IF 3.298
Yafei Duan,Yun Wang,Dalin Xiong,Jiasong Zhang
The intestine is important for nutrition, metabolism and immunity. Succinate (SA) plays a vital role in the physiological homeostasis of animal intestines. However, the effects of dietary SA on the intestinal immunity and metabolism in shrimp are not clear. In this study, we investigated the immune and metabolic responses in the intestine of Litopenaeus vannamei that were fed diets consisting of different levels of SA: 0 g/kg (Con) and 10 g/kg (SA) for 56 days. The results from a RNA-seq analysis identified 6005 differentially expressed genes (DEGs), including 2728 upregulated genes and 3277 downregulated genes, which were grouped into 312 pathways. The DEGs were most enriched in pathways related to protein synthesis and amino acid metabolism , including “ribosome”, “aminoacyl-tRNA biosynthesis”, “pyrimidine metabolism”, and “arginine and proline metabolism”; additionally, carbohydrate and lipid metabolism pathways were also activated. A large number of immune-related genes were associated with mucus barrier modification, antimicrobial activity , pathogen attachment and recognition, antioxidant activity , and apoptosis. The expression patterns of several candidate genes involved in the immune response and nutrition metabolism were detected by qPCR. This study provides insight into the transcriptomic modulating mechanisms associated with intestinal immunity and the metabolism of L. vannamei in response to the intake of dietary SA.
研究领域:
水产养殖学
·
分子生物学
·
营养学
✓ 已验证
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）