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

Properties and kinetics of microRNA regulation through canonical seed sites. microRNA通过典型种子位点调控的特性和动力学。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2015-02-16 eCollection Date: 2015-01-01

Jerry S Chen, Arra C Revilla, Michael Guerrero, Abygail M Gumbayan, Robert W Zeller

{"title":"Properties and kinetics of microRNA regulation through canonical seed sites.","authors":"Jerry S Chen, Arra C Revilla, Michael Guerrero, Abygail M Gumbayan, Robert W Zeller","doi":"","DOIUrl":"","url":null,"abstract":"MicroRNAs are a fundamental class of small RNAs involved in post-transcriptional gene regulation; however, the mechanism by which microRNAs regulate their gene targets in animals remains poorly understood. Practically, a mechanistic understanding of microRNA binding and regulation is crucial for the rational design of microRNA-based vectors for RNA interference. In this report, we focus on the largest known class of microRNA targets, the canonical seed targets, and explore the factors involved in modulating target downregulation in vivo at the protein level. Using an in vivo sensor assay in the ascidian Ciona intestinalis, we quantify miR-124-mediated downregulation of 38 canonical seed targets cloned from the Ciona genome as well as 10 control non-targets. Supporting previous findings, we observed that the seed type and number of seed sites are correlated with downregulation. However, up to a 50% variation in downregulation levels was observed for targets within the same seed class, indicating a role of non-seed factors in modulating downregulation. Although we did not observe a significant correlation of previously reported non-seed determinants with downregulation levels at saturation in our assay, our data suggest that two previously identified factors, secondary structure and 3'end complementarity, may play a role in the initial kinetics of microRNA-target binding. Importantly, using different concentrations of miR-124 we show that dose-dependent target downregulation profiles follow Michaelis-Menten kinetics. In summary, our findings emphasize the importance of non-seed factors as well as the importance of cellular concentrations of microRNAs relative to their targets when studying the mechanisms of endogenous microRNA regulation. ","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"11 ","pages":"507-14"},"PeriodicalIF":0.0,"publicationDate":"2015-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/07/30/JRGS-11-507.PMC4377283.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33089638","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

Aptamer-mediated selective delivery of short RNA therapeutics in cancer cells. 核酸适配体介导的短RNA治疗在癌细胞中的选择性递送。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2014-10-30 eCollection Date: 2014-01-01

Carla Lucia Esposito, Silvia Catuogno, Vittorio de Franciscis

{"title":"Aptamer-mediated selective delivery of short RNA therapeutics in cancer cells.","authors":"Carla Lucia Esposito, Silvia Catuogno, Vittorio de Franciscis","doi":"","DOIUrl":"","url":null,"abstract":"RNA interference (RNAi) is an important biological process that ultimately leads to suppression of gene expression. Activators of RNAi are typically small interfering RNAs (siRNA) and microRNAs (miRNA) that offer considerable therapeutic potnetial. However, a major obstacle to take these these molecules to the clinic is the absence of safe and reliable means for their specific delivery to target cells. In this regard, a highly promising class of molecules is represented by nucleic acid aptamers. These are short, structured, single-stranded RNAs or DNAs oligonucleotides that, by binding with high specificity to target molecules, provide high affinity ligands and potential antagonists of disease-associated proteins. Further, because of the high binding specificity, aptamers represent a powerful tool for the selective delivery of therapeutic cargos, including mi/siRNAs, chemotherapeutics, toxins and nanoparticles to cancer cells or tissues, thus potentially increasing the efficacy of a given therapy as well as reducing toxicity. In this review, we will focus on recent advances in the field of aptamer-mediated mi/siRNA delivery, discussing their potential and challenges in cancer therapy. ","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"10 ","pages":"500-6"},"PeriodicalIF":0.0,"publicationDate":"2014-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/98/2a/JRGS-10-500.PMC4238741.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32829261","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

Enzyme-triggered PEGylated siRNA-nanoparticles for controlled release of siRNA. 酶触发的聚乙二醇化siRNA纳米颗粒用于siRNA的控制释放。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2014-01-28 eCollection Date: 2014-01-01

Peerada Yingyuad, Mathieu Mével, Carla Prata, Christos Kontogiorgis, Maya Thanou, Andrew D Miller

{"title":"Enzyme-triggered PEGylated siRNA-nanoparticles for controlled release of siRNA.","authors":"Peerada Yingyuad, Mathieu Mével, Carla Prata, Christos Kontogiorgis, Maya Thanou, Andrew D Miller","doi":"","DOIUrl":"","url":null,"abstract":"A key goal of our recent research efforts has been to develop novel 'triggerable nanoparticle' systems with real potential utility in vivo. These are designed to be stable from the point of administration until a target site of interest is reached, then triggered for the controlled release of therapeutic agent payload(s) at the target site by changes in local endogenous conditions or through the application of some exogenous stimulus. Here we describe investigations into the use of enzymes to trigger RNAi-mediated therapy through a process of enzyme-assisted nanoparticle triggerability. Our approach is to use PEG(2000)-peptidyl lipids with peptidyl moieties sensitive to tumour-localized elastase or matrix metalloproteinase-2 digestion, and from these prepare putative enzyme-triggered PEGylated siRNA-nanoparticles. Our results provide initial proof of concept in vitro. From these data, we propose that this concept should be applicable for functional delivery of therapeutic nucleic acids to tumour cells in vivo, although the mechanism for enzyme-assisted nanoparticle triggerability remains to be fully characterized. ","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"10 ","pages":"490-9"},"PeriodicalIF":0.0,"publicationDate":"2014-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/29/61/JRGS-10-490.PMC3983657.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32269751","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

RNAi2013: RNAi at Oxford. RNAi2013:牛津的RNAi。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2013-05-20 eCollection Date: 2013-01-01

Laura A Mulcahy, David Rf Carter

引用次数: 0

Sub-cellular temporal and spatial distribution of electrotransferred LNA/DNA oligomer. 电转移LNA/DNA低聚物的亚细胞时空分布。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2013-03-15 eCollection Date: 2013-01-01

Julie Orio, Elisabeth Bellard, Houda Baaziz, Chantal Pichon, Peter Mouritzen, Marie-Pierre Rols, Justin Teissié, Muriel Golzio, Sophie Chabot

{"title":"Sub-cellular temporal and spatial distribution of electrotransferred LNA/DNA oligomer.","authors":"Julie Orio, Elisabeth Bellard, Houda Baaziz, Chantal Pichon, Peter Mouritzen, Marie-Pierre Rols, Justin Teissié, Muriel Golzio, Sophie Chabot","doi":"","DOIUrl":"","url":null,"abstract":"Low biological activity and inefficient targeted delivery in vivo have hindered RNA interference (RNAi)-based therapy from realising its full clinical potential. To overcome these hurdles, progresses have been made to develop new technologies optimizing oligonucleotides chemistry on one hand and achieving its effective delivery on the other hand. In this report, we achieved, by using the electropulsation technique (EP), efficient cellular delivery of chemically-modified oligonucleotide: The locked nucleic acid (LNA)/DNA oligomer. We used single cell level confocal fluorescence microscopy to follow the spatial and temporal distribution of electrotransferred cyanine 5 (Cy5)-labeled LNA/DNA oligomer. We observed that EP allowed LNA/DNA oligomer cellular uptake providing the oligomer a rapid access to the cytoplasm of HeLa cells. Within a few minutes after electrotransfer, Cy5-LNA/DNA oligomers shuttle from cytoplasm to nucleus whereas in absence of pulses application, Cy5-LNA/DNA oligomers were not detected. We then observed a redistribution of the Cy5 fluorescence that accumulated over time into cytoplasmic organelles. To go further and to identify these compartments, we used the HeLa GFP-Rab7 cell line to visualise late endosomes, and lysosomal or mitochondrial specific markers. Our results showed that the EP technique allowed direct entry into the cytoplasm of the Cy5-LNA/DNA oligomer bypassing the endocytosic pathway. However, in absence of pulses application, Cy5-LNA/DNA oligomer were able to enter cells through the endocytosic pathway. We demonstrated that EP is an efficient technique for LNA-based oligonucleotides delivery offering strong advantages by avoiding the endolysosomal compartmentalization, giving a rapid and free access to the cytoplasm and the nucleus where they can find their targets. ","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"9 ","pages":"479-85"},"PeriodicalIF":0.0,"publicationDate":"2013-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ad/a0/JRGS-09-479.PMC3717327.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31658244","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

Anti-PML-RARα shRNA sensitises promyelocytic leukaemia cells to all-trans retinoic acid. 抗pml - rar α shRNA使早幼粒细胞白血病细胞对全反式维甲酸敏感。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2012-01-01 Epub Date: 2012-07-13

Nicholas P Casey, Gregory M Woods

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Knockdown of AMP-activated protein kinase alpha 1 and alpha 2 catalytic subunits. amp活化的蛋白激酶α 1和α 2催化亚基的敲低。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2012-01-01 Epub Date: 2012-10-04

Larissa Tangeman, Christopher N Wyatt, Thomas L Brown

{"title":"Knockdown of AMP-activated protein kinase alpha 1 and alpha 2 catalytic subunits.","authors":"Larissa Tangeman, Christopher N Wyatt, Thomas L Brown","doi":"","DOIUrl":"","url":null,"abstract":"AMP-activated protein kinase (AMPK) is a master metabolic regulator that responds to the AMP: ATP ratio and promotes ATP production when the cell is low on energy. There are two isoforms of the catalytic alpha subunit, AMPKα1 and AMPKα2. Here, we describe the production of a small interfering RNA (siRNA) and a short hairpin RNA (shRNA) targeting both catalytic isoforms of AMPK in human, mouse, and rat. Multiple loop sequences were tested to generate the most effective shRNA. The shRNA causes significant knockdown of both isoforms of AMPKα in mouse and human cells. The shRNA effectively knocked down AMPKα1 and AMPKα2 protein levels, compared to a five basepair mismatch-control shRNA in mouse fibroblast NIH3T3 cells and significantly knocked down AMPKα1 (63%) and AMPKα2 (72%) levels compared to control in human embryonic kidney cells, HEK293s. The shRNA also causes a significant reduction in AMPK activity, measured as phosphorylation of acetyl-CoA carboxylase (ACC), a direct phosphorylation target. While the protein levels of total ACC remained the same between the AMPKα1and α2 shRNA and control shRNA-treated cells, there was a 41% reduction in phospho-ACC protein levels. The generation of this AMPKα1and α2 shRNA can be used to stably knock down protein levels and activity of both catalytic isoforms of AMPK in different species to assess function.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"8 ","pages":"470-8"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c8/d4/JRGS-08-470.PMC3542724.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31158269","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

In vivo knock-down of multidrug resistance transporters ABCC1 and ABCC2 by AAV-delivered shRNAs and by artificial miRNAs. aav递送shrna和人工mirna在体内敲除多药耐药转运体ABCC1和ABCC2

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2011-01-01 Epub Date: 2011-06-17

Florie Borel, Richard van Logtenstein, Annemart Koornneef, Piotr Maczuga, Tita Ritsema, Harald Petry, Sander Jh van Deventer, Peter Lm Jansen, Pavlina Konstantinova

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RNA silencing of lactate dehydrogenase gene in Rhizopus oryzae. 米根霉乳酸脱氢酶基因的RNA沉默。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2011-01-01 Epub Date: 2011-06-20

Ali Hashemi Gheinani, Neda Haghayegh Jahromi, Elisabeth Feuk-Lagerstedt, Mohammad J Taherzadeh

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RNAi2011: Gene Regulation by Small RNAs. RNAi2011:小rna的基因调控。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2011-01-01 Epub Date: 2011-06-01

Kate Wicks, David Rf Carter

引用次数: 0