Antisense & nucleic acid drug development最新文献_第7页

Alpha-anomeric configuration of GT oligodeoxynucleotide leads to loss of the specific aptameric and cytotoxic properties retained by the beta-anomeric analog. GT寡脱氧核苷酸的α -端粒构型导致β -端粒类似物保留的特异性核酸自适应体和细胞毒性的丧失。

Antisense & nucleic acid drug development Pub Date : 2001-12-01 DOI: 10.1089/108729001753411362

B. Scaggiante, B. Dapas, F. Quadrifoglio

引用次数: 7

Antisense oligonucleotides modulate high glucose-induced laminin overexpression and cell proliferation: a potential for therapeutic application in diabetic microangiopathy. 反义寡核苷酸调节高糖诱导的层粘连蛋白过表达和细胞增殖:糖尿病微血管病变的潜在治疗应用。

Antisense & nucleic acid drug development Pub Date : 2001-12-01 DOI: 10.1089/108729001753411353

T. Sato, G. Paryani, R. Kao, A. Li, S. Roy

{"title":"Antisense oligonucleotides modulate high glucose-induced laminin overexpression and cell proliferation: a potential for therapeutic application in diabetic microangiopathy.","authors":"T. Sato, G. Paryani, R. Kao, A. Li, S. Roy","doi":"10.1089/108729001753411353","DOIUrl":"https://doi.org/10.1089/108729001753411353","url":null,"abstract":"Vascular basement membrane (BM) thickening is a prominent and characteristic lesion of diabetic microangiopathy. Studies suggest that increased synthesis of laminin, a BM component, is associated with the development of thickened BM in diabetic vessels. In this study, we evaluated whether an interventive strategy using laminin antisense phosphorothioate oligonucleotides (Lam AS-oligos) could specifically inhibit high-glucose-induced laminin gene overexpression in vascular endothelial cells and normalize cell proliferation. Rat endothelial cells grown in high-glucose (30 mM) medium for 7 days showed increased laminin mRNA and protein level (195% +/- 28% of control, p < 0.05; 143% +/- 26% of control, p < 0.05, respectively) and reduced cell number (79% +/- 6% of control, p < 0.05) compared with cells grown in normal (5 mM) glucose medium. When cells grown in high-glucose medium were transfected with 0.4 microM Lam AS-oligos for 48 hours in the presence of 8 microM lipofectin, the laminin mRNA and protein level decreased (121% +/- 19% and 99% +/- 15% of control, respectively), and the cell number was restored to near normal level (93% +/- 7% of control). The results indicate that the antisense strategy is effective in selectively reducing laminin overexpression and improving endothelial cell proliferation under high-glucose conditions. Thus, the As-oligos may be potentially useful for preventing the development of thickened vascular BM in diabetic microangiopathy.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"33 1","pages":"387-94"},"PeriodicalIF":0.0,"publicationDate":"2001-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79261244","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: 9

Inhibition of Vesivirus infections in mammalian tissue culture with antisense morpholino oligomers. 反义morpholino低聚物对哺乳动物组织培养中Vesivirus感染的抑制作用。

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231696

D. A. Stein, D. Skilling, P. Iversen, A. W. Smith

{"title":"Inhibition of Vesivirus infections in mammalian tissue culture with antisense morpholino oligomers.","authors":"D. A. Stein, D. Skilling, P. Iversen, A. W. Smith","doi":"10.1089/108729001753231696","DOIUrl":"https://doi.org/10.1089/108729001753231696","url":null,"abstract":"Caliciviruses infect and cause disease in animals and humans. They are nonenveloped, positive-stranded RNA viruses with a genome of approximately 7.5 kb that encodes viral proteins in three open reading frames (ORF). Antisense oligomers targeting one of the three ORF of caliciviruses of the genus Vesivirus significantly inhibit viral replication in tissue culture. Porcine kidney and African green monkey kidney cells were infected with Vesivirus isolates SMSV-13 and PCV Pan-1. Phosphorodiamidate morpholino oligomers (PMO) with sequence complementary to the AUG translation start site regions of ORF1, ORF2, and ORF3 were evaluated for their effect on viral titer. Scrape-loading delivered PMO to 50%-70% of the cells of the two cell lines, as measured by fluorescence microscopy and flow cytometry. A PMO targeting ORF3 caused a significant increase in viral titer. A PMO targeting ORF2, a scrambled PMO control sequence, and an unrelated PMO antisense sequence did not alter viral titer. Various PMO sequences antisense to an upstream region of ORF1 were effective in reducing viral titer up to 80% in a dose-dependent and sequence-specific manner. The extent of viral titer reduction was proportional to the delivery of PMO to cells. These observations demonstrate that antisense PMO can disrupt caliciviral gene function in a nucleic acid sequence-specific manner and are potentially effective antiviral agents.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"26 1","pages":"317-25"},"PeriodicalIF":0.0,"publicationDate":"2001-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80156936","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: 43

Delivery of a hammerhead ribozyme specifically downregulates mutant type I collagen mRNA in a murine model of osteogenesis imperfecta. 在成骨不全小鼠模型中，锤头核酶的递送特异性下调突变型I型胶原mRNA。

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231722

I. Toudjarska, M. Kilpatrick, J. Niu, R. Wenstrup, P. Tsipouras

{"title":"Delivery of a hammerhead ribozyme specifically downregulates mutant type I collagen mRNA in a murine model of osteogenesis imperfecta.","authors":"I. Toudjarska, M. Kilpatrick, J. Niu, R. Wenstrup, P. Tsipouras","doi":"10.1089/108729001753231722","DOIUrl":"https://doi.org/10.1089/108729001753231722","url":null,"abstract":"Osteogenesis imperfecta (OI) is a systemic heritable disorder of connective tissue, caused by a mutation in one of the genes for type I collagen, whose cardinal manifestation is bone fragility. Several studies have identified two molecular mechanisms of collagen type I defects. In chain exclusion, the mutant chain is not incorporated into the collagen triple helix, whereas in chain nonexclusion, it is. The dominant-negative effect of nonexcluded mutations must be taken into account in all strategies aimed at correcting the collagen defects in individuals affected with moderate or several OI. Herein, we describe the application of hammerhead ribozymes to selectively target the mutant minigene transcript expressed in a murine calvarial osteoblast cell line. Active and control inactive ribozymes were tested in vitro on both mutant and normal targets and in the minigene-expressing cell line. Active ribozyme cleaved its target with high efficiency and specificity in both a time-dependent and dose-dependent manner. After delivery of a ribozyme expression construct, intracellular ribozyme was detected, along with a relative reduction in mutant transcript level.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"42 1","pages":"341-6"},"PeriodicalIF":0.0,"publicationDate":"2001-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83029523","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: 11

CpG motif identification for veterinary and laboratory species demonstrates that sequence recognition is highly conserved. 兽医和实验室物种的CpG基序鉴定表明序列识别是高度保守的。

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231713

R. Rankin, R. Pontarollo, X. Ioannou, A. Krieg, R. Hecker, L. Babiuk, S. van Drunen Littel-van den Hurk

引用次数: 248

High-level multiplex DNA amplification. 高水平多重DNA扩增。

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231704

N. Broude, K. Driscoll, C. Cantor

引用次数: 12

Selection and characterization of active hammerhead ribozymes targeted against cyclin E and E2F1 full-length mRNA. 针对细胞周期蛋白E和E2F1全长mRNA的活性锤头核酶的选择和鉴定。

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231669

Gabriele Grassi, Mario Grassi, J. Platz, Gerhard Bauriedel, R. Kandolf, Anne Kuhn

{"title":"Selection and characterization of active hammerhead ribozymes targeted against cyclin E and E2F1 full-length mRNA.","authors":"Gabriele Grassi, Mario Grassi, J. Platz, Gerhard Bauriedel, R. Kandolf, Anne Kuhn","doi":"10.1089/108729001753231669","DOIUrl":"https://doi.org/10.1089/108729001753231669","url":null,"abstract":"Proliferation of vascular smooth muscle cells is generally accepted as a key event in the development of restenosis following percutaneous transluminal angioplasty. To prevent human restenosis, we have designed a molecular strategy based on hammerhead ribozymes targeted against the mRNA of cyclin E and E2F1, two proteins relevant in cell cycle progression whose regulation is interconnected by a positive feedback loop. Following the identification of accessible ribozyme target sites by RNase H mapping, several hammerhead ribozymes were generated that cleave with comparable efficiency two different splice forms of cyclin E mRNA and the full-length and a truncated form of E2F1 RNA, respectively. The most active ribozymes were tested in vitro under single-turnover conditions yielding k(react)/K(m) ratios between 36 and 73 x 10(4) M(-1) min(-1), which places them in the top range ribozymes targeted against long and structured substrates. In addition, we show that the most active ribozyme selected in vitro reduces specifically and significantly (p < 0.0028) proliferation of cultured human vascular smooth muscle cells (VSMC).","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"102 1","pages":"271-87"},"PeriodicalIF":0.0,"publicationDate":"2001-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83621210","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: 11

Inhibition of GM-CSF/IL-3/IL-5 signaling by antisense oligodeoxynucleotides targeting the common beta chain of their receptors. 反义寡脱氧核苷酸对GM-CSF/IL-3/IL-5信号通路的抑制作用

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231678

M. Allam, P. Renzi

{"title":"Inhibition of GM-CSF/IL-3/IL-5 signaling by antisense oligodeoxynucleotides targeting the common beta chain of their receptors.","authors":"M. Allam, P. Renzi","doi":"10.1089/108729001753231678","DOIUrl":"https://doi.org/10.1089/108729001753231678","url":null,"abstract":"Granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-5 play a key role in allergic inflammation. They mediate their effect via receptors that consist of two distinct subunits, a cytokine-specific alpha subunit and a common beta subunit (betac) that transduces cell signaling. We sought to down-regulate the biologic activities of GM-CSF, IL-3, and IL-5 simultaneously by inhibiting betac mRNA expression with antisense technology. Experiments were performed with TF-1 cells (a human erythroleukemia cell line expressing GM-CSF, IL-3, and IL-5 receptors, which proliferates in response to these cytokines), monocytic U937 cells, which require these cytokines for differentiation, and purified human eosinophils. Cells were treated with antisense phosphorothioate oligodeoxynucleotides (ODN) targeting betac mRNA. In contrast to nontreated cells and cells treated by sense or mismatched ODN, antisense ODN inhibited betac mRNA expression and significantly decreased the level of cell surface betac protein expression on TF-1 and U937 cells. Receptor function was also affected. Antisense ODN were able to inhibit TF-1 cell proliferation in vitro in the presence of GM-CSF, IL-3, or IL-5 in the culture medium and eosinophil survival. We suggest that antisense ODN against betac may provide a new therapeutic alternative for the treatment of neoplastic or allergic diseases associated with eosinophilic inflammation.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"37 1","pages":"289-300"},"PeriodicalIF":0.0,"publicationDate":"2001-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78894030","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

Inhibition of HIV-1 replication in vitro and in human infected cells by modified antisense oligonucleotides targeting the tRNALys3/RNA initiation complex. 靶向tRNALys3/RNA起始复合物的修饰反义寡核苷酸对体外和人感染细胞中HIV-1复制的抑制作用

Antisense & nucleic acid drug development Pub Date : 2001-10-01 DOI: 10.1089/108729001753231687

F. Freund, F. Boulmé, J. Michel, M. Ventura, S. Moreau, S. Litvak

{"title":"Inhibition of HIV-1 replication in vitro and in human infected cells by modified antisense oligonucleotides targeting the tRNALys3/RNA initiation complex.","authors":"F. Freund, F. Boulmé, J. Michel, M. Ventura, S. Moreau, S. Litvak","doi":"10.1089/108729001753231687","DOIUrl":"https://doi.org/10.1089/108729001753231687","url":null,"abstract":"The untranslated 5' leader region of the human immunodeficiency virus type 1 (HIV-1) RNA plays an essential role in retroviral replication. It is the first retrotranscribed RNA region, primed from a cellular tRNALys3 partially annealed to the HIV-1 primer binding site (PBS). The structural and functional features of the HIV-1 reverse transcription initiation complex have been thoroughly studied. In this work, we used chemically modified antisense oligonucleotides (AS-ODN) as competitors of the natural tRNALys3 primer for the PBS region. Modified 2'-O-methyl AS-ODN were able to inhibit in vitro HIV-1 reverse transcription and displace the tRNALys3 previously annealed to the PBS. The destabilization of the initiation complex by 2'-O-methyl ODN was a sequence-specific process. We further demonstrated the importance of an anchor region contiguous to the PBS in the annealing of the antisense molecule, allowing the displacement of tRNALys3. The 20-mer 2'-O-methyl molecules were also able to inhibit viral replication in HIV-1-human infected cells, either by blocking cDNA synthesis during the early phase or by interfering with the annealing of the tRNALys3 primer to the PBS during the late phase of the viral cycle. Thus, the highly conserved retroviral initiation complex was shown to be a promising target when using the antisense strategy.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"1 1","pages":"301-15"},"PeriodicalIF":0.0,"publicationDate":"2001-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83096070","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: 9

CpG stimulation of primary mouse B cells is blocked by inhibitory oligodeoxyribonucleotides at a site proximal to NF-kappaB activation. CpG对原代小鼠B细胞的刺激被NF-kappaB激活位点附近的抑制性寡脱氧核糖核苷酸阻断。

Antisense & nucleic acid drug development Pub Date : 2001-08-01 DOI: 10.1089/108729001317022241

P. Lenert, L. Stunz, A. Yi, A. Krieg, R. Ashman

{"title":"CpG stimulation of primary mouse B cells is blocked by inhibitory oligodeoxyribonucleotides at a site proximal to NF-kappaB activation.","authors":"P. Lenert, L. Stunz, A. Yi, A. Krieg, R. Ashman","doi":"10.1089/108729001317022241","DOIUrl":"https://doi.org/10.1089/108729001317022241","url":null,"abstract":"Bacterial DNA and CpG-oligodeoxyribonucleotides (ODN) are powerful B cell activators, inducing apoptosis protection, cell cycle entry, proliferation, costimulatory molecule expression, immunoglobulin (Ig) and interleukin-6 (IL-6) secretion. However, proximal events in B cell activation by ODN are only partially characterized, including the translocation of NF-kappaB to the nucleus. In this paper, we provide evidence that CpG-ODN-induced cell cycle entry and apoptosis protection are blocked by SN50 or gliotoxin and thus require NF-kappaB activation. NF-kappaB activation occurred within 30 minutes of stimulation of murine B cells with a phosphorothioate (S) CpG-ODN and persisted for up to 40 hours, with p50, p65, and c-Rel as the major components. Similar to other NF-kappaB inducers, CpG-ODN caused an early IkappaBalpha and IkappaBbeta degradation plus cleavage of the p50 precursor and subsequent NF-kappaB nuclear translocation. A group of closely related S-ODN, which specifically blocked CpG-induced B cell activation at submicromolar concentrations, also prevented NF-kappaB DNA binding and transcriptional activation. These inhibitory S-ODN differed from stimulatory S-ODN by having 2-3 G substitutions in the central motif. As inhibitory S-ODN did not directly interfere with the NF-kappaB DNA binding but prevented CpG-induced NF-kappaB nuclear translocation of p50, p65, and c-Rel and blocked p105, IkappaBalpha, and IkappaBbeta degradation, we concluded that their putative target must lie upstream of inhibitory kinase (IKK) activation.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"60 1","pages":"247-56"},"PeriodicalIF":0.0,"publicationDate":"2001-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80857282","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: 119