Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research最新文献_第3页

RNAi effects on actin mRNAs in Homalodisca vitripennis cells. RNAi 对同种异形虫细胞肌动蛋白 mRNA 的影响

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2010-04-15

Cristina Rosa, Shizuo G Kamita, Haley Dequine, Ha Wuriyanghan, John A Lindbo, Bryce W Falk

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miRiam: a reliable unsupervised method for detecting miRNA binding sites on mRNAs: April 2010 release. miRiam:一种检测mrna上miRNA结合位点的可靠的无监督方法:2010年4月发布。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2010-04-01

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First siRNA library screening in hard-to-transfect HUVEC cells. 在难以转染的HUVEC细胞中首次筛选siRNA文库。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-10-29

Markus Zumbansen, Ludger M Altrogge, Nicole Ue Spottke, Sonja Spicker, Sheila M Offizier, Sandra Bs Domzalski, Allison L St Amand, Andrea Toell, Devin Leake, Herbert A Mueller-Hartmann

{"title":"First siRNA library screening in hard-to-transfect HUVEC cells.","authors":"Markus Zumbansen, Ludger M Altrogge, Nicole Ue Spottke, Sonja Spicker, Sheila M Offizier, Sandra Bs Domzalski, Allison L St Amand, Andrea Toell, Devin Leake, Herbert A Mueller-Hartmann","doi":"","DOIUrl":"","url":null,"abstract":"Meaningful RNAi-based data for target gene identification are strongly dependent on the use of a biologically relevant cell type and efficient delivery of highly functional siRNA reagents into the selected cell type. Here we report the use of the Amaxa(R) Nucleofector(R) 96-well Shuttle(R) System for siRNA screening in primary cells. Lonza's Clonetics(R) HUVEC-Human Umbilical Vein Endothelial Cells were transfected with Thermo Scientific Dharmacon siGENOME(R) siRNA Libraries targeting protein kinases and cell cycle related genes and screened for genes important for cell viability. Of the 37 primary hits, down-regulation of 33 led to reduced proliferation or increased cell death, while down-regulation of two allowed for better cell viability. The validated four genes out of the 16 strongest primary hits (COPB2, PYCS, CDK4 and MYC) influenced cell proliferation to varying degrees, reflecting differing importance for survival of HUVEC cells. Our results demonstrate that the Nucleofector(R) 96-well Shuttle(R) System allows the delivery of siRNA libraries in cell types previously considered to be difficult to transfect. Thus, identification and validation of gene targets can now be conducted in primary cells, as the selection of cell types is not limited to those accessible by lipid-mediated transfection.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"6 1","pages":"354-60"},"PeriodicalIF":0.0,"publicationDate":"2009-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/1b/35/jrgs-06-354.PMC2902142.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29121513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

'mi'-nimal interference: Somatic cell reprogramming in cancer and therapy. 动物干扰:癌症和治疗中的体细胞重编程。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-06-21

Graeme Doran

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Silencing of carbonic anhydrase in an Anopheles gambiae larval cell line, Ag55. 冈比亚按蚊幼虫细胞系Ag55中碳酸酐酶的沉默。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-06-17

Kristin E Smith, Paul J Linser

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Establishment of a positive-readout reporter system for siRNAs. sirna阳性读出报告系统的建立。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-06-12

Wei-Li Liu, Douglas P Owen, Kerry D Fisher, Leonard W Seymour, Mark Stevenson

{"title":"Establishment of a positive-readout reporter system for siRNAs.","authors":"Wei-Li Liu, Douglas P Owen, Kerry D Fisher, Leonard W Seymour, Mark Stevenson","doi":"","DOIUrl":"","url":null,"abstract":"The use of small interfering RNA molecules for therapeutic applications requires development of improved delivery systems, a process that would be facilitated by a non-invasive positive-readout mouse model for studying siRNA pharmacodynamics. Positive readout would yield better signal/noise ratios than existing negative-readout systems. We have engineered a positive-readout luciferase reporter system, activated by successful delivery of siRNA targeting the lac repressor. Co-transfection of a plasmid expressing lac repressor and a plasmid expressing firefly luciferase under the control of an RSV promoter, containing two lac operator sites, resulted in 5.7-fold lower luciferase activity than luciferase-encoding plasmid alone. Inhibition was reversed following addition of synthetic inducer, IPTG, which elevated luciferase expression to normal levels and confirmed functionality of the lac operon. Delivery of 1nM siRNA targeting lac repressor to repressor/reporter co-transfected cells was sufficient to fully restore luciferase expression to levels observed in the absence of repressor. Maximum expression was observed after 48hr, with a rapid decrease thereafter due to the short half life of luciferase. The luciferase positive-readout reporter system is therefore a dynamic indicator of successful RNAi delivery in vitro and could be adapted to generate a transgenic mouse capable of reporting RNAi activity non-invasively in vivo.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"5 1","pages":"331-8"},"PeriodicalIF":0.0,"publicationDate":"2009-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e8/91/jrgs-05-331.PMC2737236.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28416102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Non-coding RNAs: Bridging Biology and Therapy. 非编码rna:桥接生物学和治疗。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-05-22

Daniel P C Shreve, David R F Carter

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Potent subunit-specific effects on cell growth and drug sensitivity from optimised siRNA-mediated silencing of ribonucleotide reductase. 优化sirna介导的核糖核苷酸还原酶沉默对细胞生长和药物敏感性的有效亚单位特异性影响。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-03-09

Glen Reid, Natacha Coppieters 't Wallant, Rachna Patel, Ana Antonic, Faamatala Saxon-Aliifaalogo, Helen Cao, Gill Webster, James D Watson

{"title":"Potent subunit-specific effects on cell growth and drug sensitivity from optimised siRNA-mediated silencing of ribonucleotide reductase.","authors":"Glen Reid, Natacha Coppieters 't Wallant, Rachna Patel, Ana Antonic, Faamatala Saxon-Aliifaalogo, Helen Cao, Gill Webster, James D Watson","doi":"","DOIUrl":"","url":null,"abstract":"Ribonucleotide reductase (RR) has an essential role in DNA synthesis and repair and is a therapeutic target in a number of different cancers. Previous studies have shown that RNAi-mediated knockdown of either the RRM1 or RRM2 subunit sensitizes cells to the cytotoxic effects of the nucleoside analogs and more recently it has been shown that RRM2 knockdown itself has a growth inhibitory effect. Here we compare the effects of siRNA-mediated knockdown of both RRM1 and RRM2 subunits of RR in A549 and HCT-116 cells using an optimized transfection protocol. Growth of A549 cells was strongly inhibited by efficient siRNA-mediated silencing of either RRM1 or RRM2, and knockdown of each subunit led to long-term growth inhibition and cell-cycle arrest. Knockdown with sub growth inhibitory siRNA concentrations sensitized A549 and HCT-116 cells to gemcitabine when RRM1 was targeted, whereas RRM2 knockdown led to hydroxyurea sensitization. These results suggest that the inhibition of cell growth, rather than drug sensitization, is the major effect of RRM1 and RRM2 knockdown. In an A549 xenograft model, cells transfected with RRM1-specific siRNA failed to form tumors in 6 out of 8 CD1 nude mice, whereas those transfected with RRM2-specific siRNA grew but at a reduced rate. Taken together, these data demonstrate that siRNA-mediated knockdown of the RRM1 subunit is more effective than knockdown of RRM2 in inhibiting the growth of cancer cell lines and suggest that RRM1 is a potential target for nucleic acid-based cancer therapies, either alone or in combination with gemcitabine.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"5 1","pages":"321-30"},"PeriodicalIF":0.0,"publicationDate":"2009-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b0/84/jrgs-05-321.PMC2737234.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28415042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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Selective RNAi-mediated inhibition of mutated c-kit. 选择性rnai介导的突变c-kit抑制。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2009-02-20

Irene Ruano, Marta Izquierdo

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RNAi at Oxford. 牛津大学的RNAi。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2008-05-27

Masa Aleckovic, David Carter

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