Oligonucleotides最新文献_第4页

Cyclodextrin-containing polymer delivery system for light-directed siRNA gene silencing. 光导siRNA基因沉默的含环糊精聚合物递送系统。

Oligonucleotides Pub Date : 2010-08-01 DOI: 10.1089/oli.2010.0230

Sigurd Leinaes Bøe, Ane Sager Longva, Eivind Hovig

{"title":"Cyclodextrin-containing polymer delivery system for light-directed siRNA gene silencing.","authors":"Sigurd Leinaes Bøe, Ane Sager Longva, Eivind Hovig","doi":"10.1089/oli.2010.0230","DOIUrl":"https://doi.org/10.1089/oli.2010.0230","url":null,"abstract":"In this study, we have investigated the possibility of combining a cyclodextrin-containing polymer (CDP) with siRNA molecules to modulate gene expression in a light-directed manner through photochemical internalization (PCI) technology. We utilized S100A4 as a model gene to evaluate the efficacy of gene silencing. After optimization of carrier/cargo ratio and illumination dose, real-time reverse transcriptase-polymerase chain reaction data showed between 80% and 90% silencing in the siRNA samples treated with PCI compared with untreated control. In contrast, only a 0%-10% silencing effect was detected in the siRNA samples without PCI treatment, demonstrating the potency of light-specific delivery of siRNA molecules. Light-directed siRNA delivery was shown in 2 different cell lines with corresponding potency. Further, time-lapse results demonstrated maximum gene silencing only at 5 hours after endosomal release, implying, for example, rapid carrier decondensation when using the CDP. This work represents a first success in using a CDP delivery agent, without endosomolytic properties for siRNA gene silencing in a light-directed manner, opening the opportunity to use CDPs for light-directed siRNA gene silencing in vivo.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 4","pages":"175-82"},"PeriodicalIF":0.0,"publicationDate":"2010-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0230","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29133212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

Efficient gene silencing in lungs and liver using imidazole-modified chitosan as a nanocarrier for small interfering RNA. 咪唑修饰壳聚糖作为小干扰RNA纳米载体在肺和肝脏中的高效基因沉默。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2010.0235

Bilal Ghosn, Ankur Singh, Mu Li, Alexander V Vlassov, Chris Burnett, Nitin Puri, Krishnendu Roy

{"title":"Efficient gene silencing in lungs and liver using imidazole-modified chitosan as a nanocarrier for small interfering RNA.","authors":"Bilal Ghosn, Ankur Singh, Mu Li, Alexander V Vlassov, Chris Burnett, Nitin Puri, Krishnendu Roy","doi":"10.1089/oli.2010.0235","DOIUrl":"https://doi.org/10.1089/oli.2010.0235","url":null,"abstract":"Despite high specificity and potency, small interfering RNA (siRNA)-based therapeutics have been limited by their poor biostability and intracellular penetration. Thus, effective nanocarriers that can protect and efficiently deliver siRNA to target cells in vivo are needed. Here we report on the efficiency of imidazole-modified chitosan (chitosan-imidazole-4-acetic acid [IAA])-siRNA nanoparticles to mediate gene silencing after administration via either intravenous (i.v.) or intranasal (i.n.) routes. Poly(ethylene glycol) (PEG)ylated nanoparticles for i.v. delivery demonstrated significant knockdown of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) enzyme in both lung and liver at as low as 1 mg/kg siRNA dose. In addition, the efficient, dose-dependent silencing of apolipoprotein B in the liver was also shown. For i.n. delivery, significant silencing of GAPDH protein expression was seen in the lungs with only 0.5 mg/kg/day siRNA delivered over 3 consecutive days. In summary, imidazole-modified chitosan-IAA nanoparticles are potentially effective carriers for siRNA delivery.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 3","pages":"163-72"},"PeriodicalIF":0.0,"publicationDate":"2010-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0235","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29069660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 71

Physiologically based pharmacokinetics of molecular imaging nanoparticles for mRNA detection determined in tumor-bearing mice. 用于mRNA检测的分子成像纳米颗粒在荷瘤小鼠体内的生理药代动力学测定。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2009.0216

Armin W Opitz, Eric Wickstrom, Mathew L Thakur, Norman J Wagner

{"title":"Physiologically based pharmacokinetics of molecular imaging nanoparticles for mRNA detection determined in tumor-bearing mice.","authors":"Armin W Opitz, Eric Wickstrom, Mathew L Thakur, Norman J Wagner","doi":"10.1089/oli.2009.0216","DOIUrl":"https://doi.org/10.1089/oli.2009.0216","url":null,"abstract":"Disease detection and management might benefit from external imaging of disease gene mRNAs. Previously we designed molecular imaging nanoparticles (MINs) based on peptide nucleic acids complementary to cancer gene mRNAs. The MINs included contrast agents and analogs of insulin-like growth factor 1 (IGF-1). Analysis of MIN tumor uptake data showed stronger binding in tumors than in surrounding tissues. We hypothesized that MINs with an IGF-1 analog stay in circulation by binding to IGF-binding proteins. To test that hypothesis, we fit the tissue distribution results of several MINs in xenograft-bearing mice to a physiological pharmacokinetics model. Fitting experimental tissue distribution data to model-predicted mass transfer of MINs from blood into organs and tumors converged only when the parameter for MINs bound to circulating IGF-binding proteins was set to 10%-20% of the injected MIN dose. This result suggests that previous mouse imaging trials used more MINs than necessary. This prediction can be tested by a ramp of decreasing doses.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 3","pages":"117-25"},"PeriodicalIF":0.0,"publicationDate":"2010-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0216","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28934422","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: 22

Identification of RNA aptamer specific to mutant KRAS protein. KRAS蛋白突变体特异性RNA适体的鉴定。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2010.0231

Sujin Jeong, Seung Ryul Han, Young Ju Lee, Ju Hyun Kim, Seong-Wook Lee

引用次数: 21

Nucleic acid delivery with chitosan hydroxybenzotriazole. 壳聚糖羟基苯并三唑核酸递送。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2009.0227

Praneet Opanasopit, Sunee Techaarpornkul, Theerasak Rojanarata, Tanasait Ngawhirunpat, Uracha Ruktanonchai

{"title":"Nucleic acid delivery with chitosan hydroxybenzotriazole.","authors":"Praneet Opanasopit, Sunee Techaarpornkul, Theerasak Rojanarata, Tanasait Ngawhirunpat, Uracha Ruktanonchai","doi":"10.1089/oli.2009.0227","DOIUrl":"https://doi.org/10.1089/oli.2009.0227","url":null,"abstract":"The objective of this study was to investigate the transfection efficiency of chitosan hydroxybenzotriazole (CS-HOBT) for in vitro nucleic acid delivery. The results revealed that CS-HOBT was able to condense with DNA/small interfering double-stranded RNA molecules (siRNA). Illustrated by agarose gel electrophoresis, complete complexes of CS-HOBT/DNA were formed at a weight ratio of above 3, whereas those of CS-HOBT/siRNA were formed at a weight ratio of above 4 (CS molecular weights [MWs] 20 and 45 kDa) and above 2 (CS MWs 200 and 460 kDa). Gel electrophoresis results indicated that binding of CS-HOBT and DNA or siRNA depended on the MW and weight ratio. The particle sizes of CS-HOBT/nucleic acid complexes were in nanosize range. The highest transfection efficiency of CS-HOBT/DNA complex was found at a weight ratio of 2, with the lowest CS MW of 20 kDa. The CS-HOBT-mediated siRNA silencing of the enhanced green fluorescent protein gene occurred maximally with 60% efficiency. The CS-HOBT/siRNA complex with the lowest CS MW of 20 kDa at a weight ratio of 80 showed the strongest inhibition of gene expression. For cytotoxicity studies, over 80% the average cell viabilities of the complexes were observed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. This study suggests that CS-HOBT is straightforward to prepare, is safe, and exhibits significantly improved nucleic acid delivery potential in vitro.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 3","pages":"127-36"},"PeriodicalIF":0.0,"publicationDate":"2010-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0227","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28946806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 14

Topical application of antisense oligonucleotide-loaded chitosan nanoparticles to rats. 反义寡核苷酸壳聚糖纳米颗粒在大鼠体内的局部应用。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2009.0222

Suna Ozbaş-Turan, Jülide Akbuğa, Ali Demir Sezer

{"title":"Topical application of antisense oligonucleotide-loaded chitosan nanoparticles to rats.","authors":"Suna Ozbaş-Turan, Jülide Akbuğa, Ali Demir Sezer","doi":"10.1089/oli.2009.0222","DOIUrl":"https://doi.org/10.1089/oli.2009.0222","url":null,"abstract":"Skin delivery of antisense oligonucleotides (AsODNs) has exciting potential in the treatment of skin diseases. However, the therapeutic applications of oligonucleotide-based therapies are limited by the instability of these molecules toward nucleases, short half-life in vivo, and insufficient cellular uptake. The purpose of this study was to investigate in vivo antisense effect of AsODN-loaded chitosan nanoparticles after topical application. AsODN-loaded chitosan nanoparticles were topically applied to Sprague Dawley rats (adult and baby). At 1, 3, 6, 9, and 12 days posttransfection, animals' skin samples were taken for measurement of beta-galactosidase (beta-Gal) expression and histological control. After topical application of AsODN-loaded chitosan nanoparticles in different doses, beta-Gal expression reduced significantly. Highest inhibition was observed after 6 days of transfection of nanoparticles. Free AsODNs exhibited 35% of beta-Gal inhibition on the first day. beta-Gal expression was inhibited in approximately 82-85% with transfection of nanoparticles containing 30 microg AsODNs at 6 days. The antisense effect of AsODN-loaded chitosan nanoparticle in baby skin was evaluated at 6 days: 77-86% of beta-Gal suppression was measured and differences between the doses were not significant. Thus, chitosan nanoparticles are useful carrier for delivery of AsODNs into skin cells of rats and may be used for topical application on human skin.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 3","pages":"147-53"},"PeriodicalIF":0.0,"publicationDate":"2010-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0222","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28733740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 29

Dz13, a c-jun DNAzyme, is a potent inducer of caspase-2 activation. Dz13是一种c-jun DNAzyme，是一种有效的caspase-2激活诱导剂。

Oligonucleotides Pub Date : 2010-06-01 DOI: 10.1089/oli.2009.0226

Crispin R Dass, Stuart J Galloway, Peter F M Choong

{"title":"Dz13, a c-jun DNAzyme, is a potent inducer of caspase-2 activation.","authors":"Crispin R Dass, Stuart J Galloway, Peter F M Choong","doi":"10.1089/oli.2009.0226","DOIUrl":"https://doi.org/10.1089/oli.2009.0226","url":null,"abstract":"Signaling pathways for caspase-2-mediated apoptosis are poorly defined. This is partially due to a lack of a reproducible stimulus to trigger caspase-2 activation. We present the oligonucleotide Dz13, a DNA enzyme that cleaves c-Jun mRNA and is capable of inhibiting various model tumors in mice, which potently induces caspase-2 resulting in apoptosis in a panel of tumor cell lines. Dz13-mediated cell death occurred even in the absence of known caspase-2 molecular partners in p53-induced protein with a death domain, RIP-associated Ich-1/CED homologous protein with death domain, or DNA-dependent protein kinase catalytic subunit, or other caspases in cell lines of breast cancer, prostate cancer, osteosarcoma, and liposarcoma. z-VDVAD-fmk, caspase-2(-/-) mouse embryonic fibroblasts and siRNA silencing of caspase-2 in tumor cells abrogated Dz13-mediated cell death. In an orthotopic tumor model, expression of caspase-2 increased as the tumor metastasized and caspase-2 expression was sporadic in patient tumor specimens. These findings provide hope that Dz13, and other agents that evoke activation of caspase-2, may be therapeutic clinically.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 3","pages":"137-46"},"PeriodicalIF":0.0,"publicationDate":"2010-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0226","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28733662","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 17

Oligonucleotides Pub Date : 2010-04-01 DOI: 10.1089/oli.2009.0219

Crispin R Dass, Peter F M Choong

{"title":"Sequence-related off-target effect of Dz13 against human tumor cells and safety in adult and fetal mice following systemic administration.","authors":"Crispin R Dass, Peter F M Choong","doi":"10.1089/oli.2009.0219","DOIUrl":"https://doi.org/10.1089/oli.2009.0219","url":null,"abstract":"The oligonucleotide Dz13, a DNA enzyme that cleaves c-Jun mRNA, is capable of inhibiting various model tumors in mice. However, to date, a thorough examination of its target specificity in tumor cells has not been performed. In addition, an evaluation of its safety in a mammalian whole organism system has not been carried out. Dz13 mutated oligonucleotides were designed and tested in a proliferation assay. Dz13 was also tested for its safety in vivo when administered intravenously in a bolus dose, or systemically in an in utero assay. While Dz13 down-regulated target gene (c-Jun) expression in human tumor cells, c-Jun siRNA failed to cause cell growth inhibition. Furthermore, alteration of contiguous G motifs in Dz13 flanking arms inhibits cell death activity, but removal of the same from the catalytic core can increase cell death activity. A 20mer (truncated) derivative Dz13 exhibited similar activity. Dz13 was not toxic to blood and solid tissues in adult or fetal mice, though slight hepatotoxicity was noted with histology. It was also void of adverse effects to the physiological processes of angiogenesis and apoptosis. Collectively, the data support the safety of Dz13 and its activity attributed to off-target effects.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 2","pages":"51-60"},"PeriodicalIF":0.0,"publicationDate":"2010-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0219","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28707365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 14

The biological effect of an antisense oligonucleotide depends on its route of endocytosis and trafficking. 反义寡核苷酸的生物学作用取决于其内吞和转运途径。

Oligonucleotides Pub Date : 2010-04-01 DOI: 10.1089/oli.2009.0211

Md Rowshon Alam, Xin Ming, Vidula Dixit, Michael Fisher, Xiaoyuan Chen, Rudolph L Juliano

引用次数: 56

Hypo-osmotic stress enhances the uptake of polyethylenimine/oligonucleotide complexes in A549 cells via Ca(2+) mobilization from intracellular stores. 低渗透胁迫通过从细胞内储存的Ca(2+)动员增强A549细胞对聚乙烯亚胺/寡核苷酸复合物的摄取。

Oligonucleotides Pub Date : 2010-04-01 DOI: 10.1089/oli.2009.0203

Wenwu Sun, Leiting Pan, Zhuang Ma

{"title":"Hypo-osmotic stress enhances the uptake of polyethylenimine/oligonucleotide complexes in A549 cells via Ca(2+) mobilization from intracellular stores.","authors":"Wenwu Sun, Leiting Pan, Zhuang Ma","doi":"10.1089/oli.2009.0203","DOIUrl":"https://doi.org/10.1089/oli.2009.0203","url":null,"abstract":"To determine the mechanism of osmolarity involved in polyethylenimine (PEI)/oligonucleotide (ON) complex transfection in cells, we measured the fluorescence intensities of fluorescein isothiocyanate-labeled ONs complexed with PEI and the changes in cytosolic Ca(2+) concentration ([Ca(2+)](c)) in A549 cells, and we found that uptake of PEI/ON complexes was improved in the cells along with a rise of [Ca(2+)](c) in A549 cells challenged by 50% hypotonic medium. Further experiments showed that the enhanced uptake efficiency and the rise in [Ca(2+)](c) in A549 cells were almost completely abolished from cells loaded with the intracellular calcium chelator 1,2-bis(2-aminophenoxy)-N,N,N,N-tetraacetic acid-acetoxymethyl ester. 2-Aminoethoxydiphenyl borate or 8-(N,N-diethylamino) octyl-3,4,5-trimethoxybenzoate, two potent antagonists of inositol 1,4,5-trisphosphate-mediated Ca(2+) release that blunt [Ca(2+)](c) elevation via Ca(2+) release from endoplasmic reticulum, inhibited the enhanced uptake of PEI/ON complexes induced by Ca(2+)-free hypo-osmotic stress. In summary, the results strongly suggest that calcium-dependent transfection is responsible for the uptake of PEI/ON complexes into A549 cells under hypotonic conditions.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 2","pages":"111-5"},"PeriodicalIF":0.0,"publicationDate":"2010-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2009.0203","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28651676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3