Oligonucleotides最新文献_第2页

A reversible aptamer improves outcome and safety in murine models of stroke and hemorrhage. 一种可逆的适配体可改善中风和出血小鼠模型的预后和安全性。

Oligonucleotides Pub Date : 2011-02-01 Epub Date: 2010-12-13 DOI: 10.1089/oli.2010.0262

Charlene M Blake, Haichen Wang, Daniel T Laskowitz, Bruce A Sullenger

{"title":"A reversible aptamer improves outcome and safety in murine models of stroke and hemorrhage.","authors":"Charlene M Blake, Haichen Wang, Daniel T Laskowitz, Bruce A Sullenger","doi":"10.1089/oli.2010.0262","DOIUrl":"10.1089/oli.2010.0262","url":null,"abstract":"Treatment of acute ischemic stroke with intravenous tissue-type plasminogen activator is underutilized partly due to the risk of life-threatening hemorrhage. In response to the clinical need for safer stroke therapy, we explored using an aptamer-based therapeutic strategy to promote cerebral reperfusion in a murine model of ischemic stroke. Aptamers are nucleic acid ligands that bind to their targets with high affinity and specificity, and can be rapidly reversed with an antidote. Here we show that a Factor IXa aptamer administered intravenously after 60 minutes of cerebral ischemia and reperfusion improved neurological function and was associated with reduced thrombin generation and decreased inflammation. Moreover, when the aptamer was administered in the setting of intracranial hemorrhage, treatment with its specific antidote reduced hematoma volume and improved survival. The ability to rapidly reverse a pharmacologic agent that improves neurological function after ischemic stroke should intracranial hemorrhage arise indicates that aptamer-antidote pairs may represent a novel, safer approach to treatment of stroke.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"21 1","pages":"11-9"},"PeriodicalIF":0.0,"publicationDate":"2011-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3043993/pdf/oli.2010.0262.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29522573","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

Efficiency of cellular delivery of antisense peptide nucleic acid by electroporation depends on charge and electroporation geometry. 细胞电穿孔传递反义肽核酸的效率取决于电荷和电穿孔几何。

Oligonucleotides Pub Date : 2011-02-01 Epub Date: 2011-01-14 DOI: 10.1089/oli.2010.0266

Mette Joergensen, Birgit Agerholm-Larsen, Peter E Nielsen, Julie Gehl

{"title":"Efficiency of cellular delivery of antisense peptide nucleic acid by electroporation depends on charge and electroporation geometry.","authors":"Mette Joergensen, Birgit Agerholm-Larsen, Peter E Nielsen, Julie Gehl","doi":"10.1089/oli.2010.0266","DOIUrl":"https://doi.org/10.1089/oli.2010.0266","url":null,"abstract":"Electroporation is potentially a very powerful technique for both in vitro cellular and in vivo drug delivery, particularly relating to oligonucleotides and their analogs for genetic therapy. Using a sensitive and quantitative HeLa cell luciferase RNA interference mRNA splice correction assay with a functional luciferase readout, we demonstrate that parameters such as peptide nucleic acid (PNA) charge and the method of electroporation have dramatic influence on the efficiency of productive delivery. In a suspended cell electroporation system (cuvettes), a positively charged PNA (+8) was most efficiently transferred, whereas charge neutral PNA was more effective in a microtiter plate electrotransfer system for monolayer cells. Surprisingly, a negatively charged (-23) PNA did not show appreciable activity in either system. Findings from the functional assay were corroborated by pulse parameter variations, polymerase chain reaction, and confocal microscopy. In conclusion, we have found that the charge of PNA and electroporation system combination greatly influences the transfer efficiency, thereby illustrating the complexity of the electroporation mechanism.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"21 1","pages":"29-37"},"PeriodicalIF":0.0,"publicationDate":"2011-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0266","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29598271","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: 25

Effect of G-rich oligonucleotides on the proliferation of leukemia cells and its relationship with p53 expression. 富含 G 的寡核苷酸对白血病细胞增殖的影响及其与 p53 表达的关系。

Oligonucleotides Pub Date : 2011-02-01 Epub Date: 2011-01-19 DOI: 10.1089/oli.2010.0254

Lei Zhi, Jianwei Zhang, Yujiao Jia, Shilong Shan, Yan Li, Donghai Wang, Min Wang, Qing Rao, Haiyan Xing, Kejing Tang, Zheng Tian, Jianxiang Wang, Yingchang Mi

{"title":"Effect of G-rich oligonucleotides on the proliferation of leukemia cells and its relationship with p53 expression.","authors":"Lei Zhi, Jianwei Zhang, Yujiao Jia, Shilong Shan, Yan Li, Donghai Wang, Min Wang, Qing Rao, Haiyan Xing, Kejing Tang, Zheng Tian, Jianxiang Wang, Yingchang Mi","doi":"10.1089/oli.2010.0254","DOIUrl":"10.1089/oli.2010.0254","url":null,"abstract":"G-rich oligonucleotides (GROs) can inhibit cell proliferation by inducing cell cycle arrest at S phase in tumor cell lines. GROs bind specific cellular proteins, such as nucleolin, a crucial protein interacting with P53; however, little is known about the relationship between GROs and P53. In this study, we have shown that GROs inhibited the proliferation of U937 cells (a human monocytic leukemia cell line without P53 expression) by inducing S-phase arrest. We also showed that GRO colocalized with nucleolin in U937 cells. GRO treatment induced alteration of a series of cell cycle regulatory proteins in U937 cells. Increased Cdk2 expression might promote the cells to enter S phase and subsequent decrease of Cdk2 might induce cell cycle arrest in S phase. Transfection of U937 cells with a wild-type p53 gene caused the formation of nucleolin-P53 complex, which alleviated the effect of GRO on leukemia cells. This alleviated effect is probably due to the decreased uptake of GRO.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"21 1","pages":"21-7"},"PeriodicalIF":0.0,"publicationDate":"2011-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0254","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29610306","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: 5

The RNA activator ds-p21 potentiates the cytotoxicity induced by fludarabine in Dohh2 cells. RNA激活剂ds-p21增强了氟达拉滨对doh2细胞的细胞毒性。

Oligonucleotides Pub Date : 2011-02-01 Epub Date: 2011-01-17 DOI: 10.1089/oli.2010.0249

Francesca Patella, Serena Lucotti, Milena Rizzo, Monica Evangelista, Giuseppe Rainaldi

{"title":"The RNA activator ds-p21 potentiates the cytotoxicity induced by fludarabine in Dohh2 cells.","authors":"Francesca Patella, Serena Lucotti, Milena Rizzo, Monica Evangelista, Giuseppe Rainaldi","doi":"10.1089/oli.2010.0249","DOIUrl":"https://doi.org/10.1089/oli.2010.0249","url":null,"abstract":"Recently, it has been reported that, in several tumor cell lines, short double-stranded RNAs tailored for promoter regions of specific genes are able to activate their transcription. Such molecules (named RNA activators) act opposite to other double-stranded RNA molecules (named RNA inhibitors) in that the overexpression instead of underexpression of a given gene is triggered. In Dohh2 non-Hodgkin lymphoma cells, the transcriptional repressor BCL6, which negatively controls both p53 and p21, is overexpressed, so that the cells can escape the check point governed by p53 and proliferate. The aim of this work was to investigate whether the RNA activator p21 can represent a tool to circumvent the transcriptional control of BCL6 and induce the blockage of cell proliferation in Dohh2 non-Hodgkin lymphoma cells. For that, Dohh2 cells were transfected with either a control RNA activator (ds-NC) or an RNA activator specific for human p21 promoter (ds-p21). At various time points after transfection, the cells were collected and p21 was measured. Dohh2 cells transfected with ds-p21 showed a slight but significant overexpression of p21 at both mRNA and protein levels. Nonetheless, cell proliferation, cell cycle, and apoptosis were not significantly modified. In contrast, the exposure of Dohh2 cells transfected with ds-p21 to fludarabine potentiates the cytotoxicity of the drug, suggesting the RNA activator p21 complements the fludarabine-dependent cell death pathways.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"21 1","pages":"39-45"},"PeriodicalIF":0.0,"publicationDate":"2011-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0249","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29604775","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

Alan M. Gewirtz, M.D. Alan M. Gewirtz医学博士

Oligonucleotides Pub Date : 2010-12-18 DOI: 10.1089/OLI.2010.1500

C. Abrams

引用次数: 0

In vitro metabolic stabilities and metabolism of 2'-O-(methoxyethyl) partially modified phosphorothioate antisense oligonucleotides in preincubated rat or human whole liver homogenates. 2'- o -(甲氧基乙基)部分修饰的硫代磷酸酯反义寡核苷酸在预孵育大鼠或人全肝匀浆中的体外代谢稳定性和代谢。

Oligonucleotides Pub Date : 2010-12-01 Epub Date: 2010-11-30 DOI: 10.1089/oli.2010.0252

Min-Son Baek, Rosie Z Yu, Hans Gaus, John S Grundy, Richard S Geary

{"title":"In vitro metabolic stabilities and metabolism of 2'-O-(methoxyethyl) partially modified phosphorothioate antisense oligonucleotides in preincubated rat or human whole liver homogenates.","authors":"Min-Son Baek, Rosie Z Yu, Hans Gaus, John S Grundy, Richard S Geary","doi":"10.1089/oli.2010.0252","DOIUrl":"https://doi.org/10.1089/oli.2010.0252","url":null,"abstract":"In vitro metabolic stability testing of phosphorothioate 2'-O-methoxyethyl (2'-MOE) partially modified antisense oligonucleotides (ASOs) is not routinely performed to help screen discovery compounds (eg, predict in vivo half-lives), as no suitable in vitro test system currently exists. The aims of this work were to develop, optimize, and evaluate an in vitro whole liver homogenate (rat or human) test system. The test system was used to evaluate in vitro metabolic stabilities (intrinsic clearance) of selected ASOs, with results compared to reported in vivo half-lives, and generated metabolites also identified. Test system optimization involved preincubating whole liver homogenates at 37°C for ≥24 hours, which increased in vitro ASO metabolism rate. From calculated in vitro intrinsic clearance (CL(int)) values in preincubated rat or human whole liver homogenates, metabolic stabilities of fully phosphorothioated 2'-MOE ASOs (ISIS 104838 and ISIS 301012) were, as expected, greater (ie, lower CL(int)) than a 2'-MOE ASO containing a single phosphodiester substitution (ISIS 104838PO10). However, comparable-to-lower in vitro metabolic stability for ISIS 301012 was seen compared to ISIS 104838, in contrast to reported ∼2-fold longer in vivo tissue elimination half-lives for ISIS 301012. Identified in vitro metabolic products of ISIS 301012 were consistent with previously reported in vivo observations.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 6","pages":"309-16"},"PeriodicalIF":0.0,"publicationDate":"2010-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0252","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29501191","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: 31

Suppression of hepatitis C virus genome replication in cells with RNA-cleaving DNA enzymes and short-hairpin RNA. RNA切割DNA酶和短发夹RNA抑制丙型肝炎病毒基因组在细胞中的复制。

Oligonucleotides Pub Date : 2010-12-01 Epub Date: 2010-09-23 DOI: 10.1089/oli.2010.0256

Bokhui Lee, Kyung Bo Kim, Sangtaek Oh, Joon Sig Choi, Jong-Sang Park, Dal-Hee Min, Dong-Eun Kim

{"title":"Suppression of hepatitis C virus genome replication in cells with RNA-cleaving DNA enzymes and short-hairpin RNA.","authors":"Bokhui Lee, Kyung Bo Kim, Sangtaek Oh, Joon Sig Choi, Jong-Sang Park, Dal-Hee Min, Dong-Eun Kim","doi":"10.1089/oli.2010.0256","DOIUrl":"https://doi.org/10.1089/oli.2010.0256","url":null,"abstract":"A class of antisense oligodeoxyribozymes, known as the 10-23 DNA enzymes (DNAzyme), has been shown to efficiently cleave target RNA at purine-pyrimidine junctions in vitro. Herein we have utilized a strategy to identify accessible cleavage sites for DNAzyme in the target RNA, the hepatitis C virus nonstructural gene 3 (HCV NS3) RNA that encodes viral helicase and protease, from a pool of randomized DNAzyme library. The screening procedure identified 18 potential cleavage sites in the target RNA. Corresponding DNAzymes were constructed for the selected target sites and were tested for RNA cleavage in vitro. Using positively charged dendrimer nanoparticles, the target RNA-cleaving DNAzymes that are 31-mer oliogonucleotides are delivered into the human hepatoma cells harboring the HCV subgenomic replicon RNA. DNAzymes introduced into the cells efficiently inhibited HCV RNA replication by reducing the expression of HCV NS3. In addition, we designed short-hairpin RNA (shRNA) that targets the same cleavage site for the selected DNAzyme and confirmed that the shRNA also inhibited HCV NS3 gene expression in the HCV replicon cells. These selected DNAzyme and shRNA may be a viable therapeutic intervention to inhibit HCV replication in hepatic cells. We suggest that the method used in this study can be applicable for identification of available sites in any target RNA for antisense oligonucleotides and siRNAs.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 6","pages":"285-96"},"PeriodicalIF":0.0,"publicationDate":"2010-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0256","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29297657","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: 13

Pitfalls of cell-systematic evolution of ligands by exponential enrichment (SELEX): existing dead cells during in vitro selection anticipate the enrichment of specific aptamers. 通过指数富集(SELEX)进行配体的细胞系统进化的陷阱:在体外选择过程中存在的死细胞预期特定适配体的富集。

Oligonucleotides Pub Date : 2010-12-01 Epub Date: 2010-12-06 DOI: 10.1089/oli.2010.0253

Meltem Avci-Adali, Markus Metzger, Nadja Perle, Gerhard Ziemer, Hans P Wendel

{"title":"Pitfalls of cell-systematic evolution of ligands by exponential enrichment (SELEX): existing dead cells during in vitro selection anticipate the enrichment of specific aptamers.","authors":"Meltem Avci-Adali, Markus Metzger, Nadja Perle, Gerhard Ziemer, Hans P Wendel","doi":"10.1089/oli.2010.0253","DOIUrl":"https://doi.org/10.1089/oli.2010.0253","url":null,"abstract":"Aptamers represent auspicious ligands for recognition of target molecules on the surface of a specific cell population, such as stem or cancer cells. These ligands are able to capture and enrich desired cells from a cell mixture, and can be used for identification of new biomarkers, development of cell-specific therapeutics, and stem cell therapy. In this study, we investigated the influence of dead cells on single-stranded DNA (ssDNA) binding and established a method to eliminate dead cells from a cell suspension. Flow cytometry analyses demonstrated that all dead cells were stained with fluorescein-labeled ssDNA molecules. The increasing of the proportion of dead cells led to an increased number of cells that were positive for ssDNA staining. Using dead cell removal microbeads, the proportion of dead cells was significantly reduced. The studies demonstrated that dead cells lead to unspecific uptake/binding of ssDNA molecules during cell-Systematic Evolution of Ligands by Exponential enrichment (SELEX) and can cause failure of the selection process. Thus, the elimination of dead cell population before incubation with ssDNA molecules will reduce the loss of target binding sequences and the contamination of the enriched aptamer pool with unspecific ssDNA molecules caused by unspecific binding to dead cells.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 6","pages":"317-23"},"PeriodicalIF":0.0,"publicationDate":"2010-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0253","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29515070","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: 41

Site-specific DNA photocleavage and photomodulation by oligonucleotide conjugates. 寡核苷酸偶联物的位点特异性DNA光切割和光调节。

Oligonucleotides Pub Date : 2010-12-01 Epub Date: 2010-09-23 DOI: 10.1089/oli.2010.0247

Netanel Kolevzon, Eylon Yavin

引用次数: 6

Intracellular traffic of oligodeoxynucleotides in and out of the nucleus: effect of exportins and DNA structure. 细胞核内外寡脱氧核苷酸的细胞内运输:输出蛋白和DNA结构的影响。

Oligonucleotides Pub Date : 2010-12-01 Epub Date: 2010-10-14 DOI: 10.1089/oli.2010.0255

Stephen J Forsha, Irina V Panyutin, Ronald D Neumann, Igor G Panyutin

{"title":"Intracellular traffic of oligodeoxynucleotides in and out of the nucleus: effect of exportins and DNA structure.","authors":"Stephen J Forsha, Irina V Panyutin, Ronald D Neumann, Igor G Panyutin","doi":"10.1089/oli.2010.0255","DOIUrl":"https://doi.org/10.1089/oli.2010.0255","url":null,"abstract":"The delivery of oligodeoxynucleotides (ODNs) into cells is widely utilized for antisense, antigene, aptamer, and similar approaches to regulate gene and protein activities based upon the ODNs' sequence-specific recognition. Short pieces of DNA can also be generated in biological processes, for example, after degradation of viral or bacterial DNA. However, the mechanisms that regulate intracellular trafficking and localization of ODNs are not fully understood. Here we study the effects of major transporters of microRNA, exportin-1 (Exp1) and exportin-5 (Exp5), on the transport of single-stranded ODNs in and out of the nucleus. For this, we employed a fluorescent microscopy-based assay to quantitatively measure the redistribution of ODNs between the nucleus and cytoplasm of live cells. By measuring the fluorescent signal of the nuclei we observed that after delivery into cells via cationic liposomes ODNs rapidly accumulated inside nuclei. However, after removal of the ODN/liposome containing media, we found re-localization of ODNs from the nuclei to cytoplasm of the cells over the time course of several hours. Downregulation of the Exp5 gene by siRNA resulted in a slight increase of ODN uptake into the nucleus, but the kinetics of ODN efflux to the cytoplasm was not affected. Inhibition of Exp1 with leptomycin B somewhat slowed down the clearance of ODNs from the nucleus; however, within 6 hours most of the ODN were still being cleared form the nucleus. ODNs that could form intramolecular G-quadruplex structures behaved differently. They also accumulated in nuclei, although at a lesser extent than unstructured ODN, but they remained there for up to 20 hours after transfection, causing significant cell death. We conclude that Exp1 and Exp5 are not the major transporters of our ODNs out of the nucleus, and that the transport of ODNs is strongly affected by their secondary structure.","PeriodicalId":19523,"journal":{"name":"Oligonucleotides","volume":"20 6","pages":"277-84"},"PeriodicalIF":0.0,"publicationDate":"2010-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/oli.2010.0255","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29352361","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: 10