• Title/Summary/Keyword: Interference regulation

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New Hairpin RNAi Vector with Brassica rapa ssp. pekinensis Intron for Gene Silencing in Plants

  • Lee, Gi-Ho;Lee, Gang-Seob;Park, Young-Doo
    • 원예과학기술지
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    • 제35권3호
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    • pp.323-332
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    • 2017
  • Homology-specific transcriptional and post-transcriptional silencing, an intrinsic mechanism of gene regulation in most eukaryotes, can be induced by anti-sense, co-suppression, or hairpin-based double-stranded RNA. Hairpin-based RNA interference (RNAi) has been applied to analyze gene function and genetically modify crops. However, RNAi vector construction usually requires high-cost cloning steps and large amounts of time, or involves methods that are protected by intellectual property rights. We describe a more effective method for generating intron-spliced RNAi constructs. To produce intron-spliced hairpin RNA, an RNAi cassette was ligated with the first intron and splicing sequences of the Brassica rapa ssp. pekinensis histone deacetylase 1 gene. This method requires a single ligation of the PCR-amplified target gene to SpeI-NcoI and SacI-BglII enzyme sites to create a gene-specific silencing construct. We named the resulting binary vector system pKHi and verified its functionality by constructing a vector to silence DIHYDROFLAVONOL 4-REDUCTASE (DFR), transforming it into tobacco plants, and confirming DFR gene-silencing via PCR, RT-qPCR, and analysis of the accumulation of small interfering RNAs. Reduction of anthocyanin biosynthesis was also confirmed by analyzing flower color of the transgenic tobacco plants. This study demonstrates that small interfering RNAs generated through the pKHi vector system can efficiently silence target genes and could be used in developing genetically modified crops.

Caffeine Induces the Stress Response and Up-Regulates Heat Shock Proteins in Caenorhabditis elegans

  • Al-Amin, Mohammad;Kawasaki, Ichiro;Gong, Joomi;Shim, Yhong-Hee
    • Molecules and Cells
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    • 제39권2호
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    • pp.163-168
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    • 2016
  • Caffeine has both positive and negative effects on physiological functions in a dose-dependent manner. C. elegans has been used as an animal model to investigate the effects of caffeine on development. Caffeine treatment at a high dose (30 mM) showed detrimental effects and caused early larval arrest. We performed a comparative proteomic analysis to investigate the mode of action of high-dose caffeine treatment in C. elegans and found that the stress response proteins, heat shock protein (HSP)-4 (endoplasmic reticulum [ER] chaperone), HSP-6 (mitochondrial chaperone), and HSP-16 (cytosolic chaperone), were induced and their expression was regulated at the transcriptional level. These findings suggest that high-dose caffeine intake causes a strong stress response and activates all three stress-response pathways in the worms, including the ER-, mitochondrial-, and cytosolic pathways. RNA interference of each hsp gene or in triple combination retarded growth. In addition, caffeine treatment stimulated a food-avoidance behavior (aversion phenotype), which was enhanced by RNAi depletion of the hsp-4 gene. Therefore, up-regulation of hsp genes after caffeine treatment appeared to be the major responses to alleviate stress and protect against developmental arrest.

Transcriptional Profiling and Dynamical Regulation Analysis Identify Potential Kernel Target Genes of SCYL1-BP1 in HEK293T Cells

  • Wang, Yang;Chen, Xiaomei;Chen, Xiaojing;Chen, Qilong;Huo, Keke
    • Molecules and Cells
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    • 제37권9호
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    • pp.691-698
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    • 2014
  • SCYL1-BP1 is thought to function in the p53 pathway through Mdm2 and hPirh2, and mutations in SCYL1-BP1 are associated with premature aging syndromes such as Geroderma Osteodysplasticum; however, these mechanisms are unclear. Here, we report significant alterations in miRNA expression levels when SCYL1-BP1 expression was inhibited by RNA interference in HEK293T cells. We functionally characterized the effects of potential kernel miRNA-target genes by miRNA-target network and protein-protein interaction network analysis. Importantly, we showed the diminished SCYL1-BP1 dramatically reduced the expression levels of EEA1, BMPR2 and BRCA2 in HEK293T cells. Thus, we infer that SCYL1-BP1 plays a critical function in HEK293T cell development and directly regulates miRNA-target genes, including, but not limited to, EEA1, BMPR2, and BRCA2, suggesting a new strategy for investigating the molecular mechanism of SCYL1-BP1.

An Antioxidative and Antiinflammatory Agent for Potential Treatment of Osteoarthritis from Ecklonia cava

  • Shin Hyeon-Cheol;Hwang Hye Jeong;Kang Kee Jung;Lee Bong Ho
    • Archives of Pharmacal Research
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    • 제29권2호
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    • pp.165-171
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    • 2006
  • Osteoarthritis is thought to be induced by the ageing-related loss of homeostatic balance between degeneration and repair mechanism around cartilage tissue in which inflammatory mediators such as reactive oxygen species, cytokines and prostaglandins are prone to overproduction under undesirable physiological conditions. Phlorotannins are unique polyphenolic compounds bearing dibenzo-1,4-dioxin skeleton which are not found in terrestrial plants but found only in some brown algal species such as Ecklonia and Eisenia families. Phlorotanninrich extracts of Ecklonia cava including LAD103 showed significant antioxidant activities such as DPPH radical scavenging, ferric ion reduction, peroxynitrite scavenging, and inhibition of LDL oxidation, indicating their possible antioxidative interference both in onset and downstream consequences of osteoarthritis. LAD103 also showed significant down regulation of $PGE_2$ generation in LPS-treated RAW 246.7 cells, and significant inhibition of human recombinant interleukin-$1{\alpha}$-induced proteoglycan degradation, indicating its beneficial involvement in pathophysiological consequences of osteoarthritis, the mechanism of which needs further investigation. Since LAD103 showed strong therapeutic potentials in arthritic treatment through several in vitro experiments, it is highly encouraged to perform further mechanistic and efficacy studies.

개발사업지구 내 자연생태계 가치평가모델 개발 및 적용 (Development and Application of Natural Ecosystem Value Assessment Model in Land Development)

  • 이동근;이현이;김은영;최재용
    • 한국환경복원기술학회지
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    • 제12권3호
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    • pp.59-71
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    • 2009
  • The goals of this study were to development Natural Ecosystem Value Assessment Model, to suggest the practical applicability of the Natural Ecosystem value assessment for development areas, and to assess natural ecosystem quantitatively using GIS. The Model can evaluate Natural Ecosystem Value between before and after land developments. To select indicators for the model, we conducted the literature reviews. and six indicators and the standard of evaluation had been developed through the expert interviews and literature reviews. the relative importance of the evaluation index throught two times expert questionnaires was found out. the Natural Ecosystem Value Assessment Model has been established, The results demonstrate that although the suggested six indicators are arranged by priority, to maintain quantitative and qualitative the natural ecosystem value, all indicators are desirable to be maintained complementarily. The findings of this study suggest that the natural ecosystem value assessment model appears to be effective an assessment for damaged natural environment value and possible to assign a score value. Also, this model can be applied to research areas and has implications to help maintain the natural ecosystem in land development districts.

선박보안경보장치(Ship Security Alert System)의 기능 및 기술적 특성 연구 (A Study on Technical features and characteristics for Ship Security Alert Systems)

  • 장동원
    • 한국정보통신학회:학술대회논문집
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    • 한국해양정보통신학회 2004년도 춘계종합학술대회
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    • pp.85-90
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    • 2004
  • 선박보안경보장치(Ship Security Alert System)는 2001년 미국 9월11일 테러 이후에 전 세계적으로 이에 대응하기 위한 방법으로 해상에서 선박을 통해 발생될 수 있는 테러에 대비하기 위한 통신시스템으로 2004년 7월1일 이후에 건조된 선박에는 반드시 탑재되어야 한다. 이 장치는 IMO SOLAS 국제 조약에 따르며 선박의 안전이 위협을 받고 있거나 위태롭게 되었음을 해당기관에 표시하기 위하여 육상으로 보안경보를 송신할 목적으로 선박에 설치되어야 한다. 무선통신설비는 특별히 정하지는 않았지만 전 세계를 포함하기 위하여 위성망을 사용하는 것이 유리하다. 본 고에서는 선박보안경보장치의 필수 기능 및 사용 가능한 통신시스템 특성에 대해서 분석하였으며 국내 관련 기술기준 방향에 대하여 제안하였다.

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Differential Game Theoretic Approach for Distributed Dynamic Cooperative Power Control in Cognitive Radio Ad Hoc Networks

  • Zhang, Long;Huang, Wei;Wu, Qiwu;Cao, Wenjing
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • 제9권10호
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    • pp.3810-3830
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    • 2015
  • In this paper, we investigate the differential game theoretic approach for distributed dynamic cooperative power control in cognitive radio ad hoc networks (CRANETs). First, a payoff function is defined by taking into consideration the tradeoff between the stock of accumulated power interference to the primary networks and the dynamic regulation of the transmit power of secondary users (SUs). Specifically, the payoff function not only reflects the tradeoff between the requirement for quickly finding the stable available spectrum opportunities and the need for better channel conditions, but also reveals the impact of the differentiated types of data traffic on the demand of transmission quality. Then the dynamic power control problem is modeled as a differential game model. Moreover, we convert the differential game model into a dynamic programming problem to obtain a set of optimal strategies of SUs under the condition of the grand coalition. A distributed dynamic cooperative power control algorithm is developed to dynamically adjust the transmit power of SUs under grand coalition. Finally, numerical results are presented to demonstrate the effectiveness of the proposed algorithm for efficient power control in CRANETs.

Silencing YY1 Alleviates Ox-LDL-Induced Inflammation and Lipid Accumulation in Macrophages through Regulation of PCSK9/ LDLR Signaling

  • Zhengyao Qian;Jianping Zhao
    • Journal of Microbiology and Biotechnology
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    • 제32권11호
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    • pp.1406-1415
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    • 2022
  • The formation of macrophage foam cells stimulated by oxidized low-density lipoprotein (ox-LDL) is deemed an important cause of atherosclerosis. Transcription factor Yin Yang 1 (YY1), which is a universally expressed multifunctional protein, is closely related to cell metabolism disorders such as lipid metabolism, sugar metabolism, and bile acid metabolism. However, whether YY1 is involved in macrophage inflammation and lipid accumulation still remains unknown. After mouse macrophage cell line RAW264.7 cells were induced by ox-LDL, YY1 and proprotein convertase subtilisin/kexin type 9 (PCSK9) expressions were found to be increased while low-density lipoprotein receptor (LDLR) expression was lowly expressed. Subsequently, through reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot analysis, Oil Red O staining and cholesterol quantification, it turned out that silencing of YY1 attenuated the inflammatory response and lipid accumulation in RAW264.7 cells caused by ox-LDL. Moreover, results from the JASPAR database, chromatin immunoprecipitation (ChIP) assay, luciferase reporter assay and Western blot analysis suggested that YY1 activated PCSK9 by binding to PCSK9 promoter and modulated the expression of LDLR in the downstream of PCSK9. In addition, the results of functional experiments demonstrated that the inhibitory effects of YY1 interference on ox-LDL-mediated macrophage inflammation and lipid accumulation were reversed by PCSK9 overexpression. To sum up, YY1 depletion inhibited its activation of PCSK9, thereby reducing cellular inflammatory response, cholesterol homeostasis imbalance, and lipid accumulation caused by ox-LDL.

SAFB1, an RBMX-binding protein, is a newly identified regulator of hepatic SREBP-1c gene

  • Omura, Yasushi;Nishio, Yoshihiko;Takemoto, Tadashi;Ikeuchi, Chikako;Sekine, Osamu;Morino, Katsutaro;Maeno, Yasuhiro;Obata, Toshiyuki;Ugi, Satoshi;Maegawa, Hiroshi;Kimura, Hiroshi;Kashiwagi, Atsunori
    • BMB Reports
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    • 제42권4호
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    • pp.232-237
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    • 2009
  • Sterol regulatory element-binding protein (SREBP)-1c plays a crucial role in the regulation of lipogenic enzymes in the liver. We previously reported that an X-chromosome-linked RNA binding motif (RBMX) regulates the promoter activity of Srebp-1c. However, still unknown was how it regulates the gene expression. To elucidate this mechanism, we screened the cDNA library from mouse liver by yeast two-hybrid assay using RBMX as bait and identified scaffold attachment factor B1 (SAFB1). Immunoprecipitation assay demonstrated binding of SAFB1 to RBMX. Chromatin immunoprecipitation assay showed binding of both SAFB1 and RBMX to the upstream region of Srebp-1c gene. RNA interference of Safb1 reduced the basal and RBMX-induced Srebp-1c promoter activities, resulting in reduced Srebp-1c gene expression. The effect of SAFB1 overexpression on Srebp-1c promoter was found only in the presence of RBMX. These results indicate a major role for SAFB1 in the activation of Srebp-1c through its interaction with RBMX.