Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research最新文献_第6页

Manipulating and enhancing the RNAi response. 操纵和增强RNAi反应。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2006-01-12

Peter I Joyce, Joseph M Gallagher, Patricia E Kuwabara

引用次数: 0

P-bodies and RNAi: The missing link? P-体和RNAi：缺失的环节？

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-12-16

Derek M Dykxhoorn

引用次数: 0

Designer siRNAs to overcome the challenges from the RNAi pathway. 设计sirna以克服来自RNAi途径的挑战。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-11-30

Sumedha D Jayasena

引用次数: 0

A novel approach for inhibition of HIV-1 by RNA interference: counteracting viral escape with a second generation of siRNAs. 通过RNA干扰抑制HIV-1的新方法:用第二代sirna对抗病毒逃逸。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-10-14

Olivier ter Brake, Ben Berkhout

{"title":"A novel approach for inhibition of HIV-1 by RNA interference: counteracting viral escape with a second generation of siRNAs.","authors":"Olivier ter Brake, Ben Berkhout","doi":"","DOIUrl":"","url":null,"abstract":"RNA interference (RNAi) is an evolutionary conserved gene silencing mechanism in which small interfering RNA (siRNA) mediates the sequence specific degradation of mRNA. The recent discovery that exogenously delivered siRNA can trigger RNAi in mammalian cells raises the possibility to use this technology as a therapeutic tool against pathogenic viruses. Indeed, it has been shown that siRNAs can be used effectively to inhibit virus replication. The focus of this review is on RNA interference strategies against HIV-1 and how this new technology may be developed into a new successful therapy. One of the hallmarks of RNAi, its sequence specificity, also presents a way out for the virus, as single nucleotide substitutions in the target region can abolish the suppression. Strategies to prevent the emergence of resistant viruses have been suggested and involve the targeting of conserved sequences and the simultaneous use of multiple siRNAs, similar to current highly active antiretroviral therapy. We present an additional strategy aimed at preventing viral escape by using a second generation of siRNAs that recognize the mutated target sites.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"1 2","pages":"56-65"},"PeriodicalIF":0.0,"publicationDate":"2005-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737200/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28416437","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

RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA. 通过转染小干扰RNA的二聚体和三聚体在哺乳动物和果蝇细胞中诱导RNAi。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-10-14

Kumiko Ui-Tei, Shuhei Zenno, Aya Juni, Kaoru Saigo

{"title":"RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA.","authors":"Kumiko Ui-Tei, Shuhei Zenno, Aya Juni, Kaoru Saigo","doi":"","DOIUrl":"","url":null,"abstract":"Twenty one base pair long small interfering RNAs (siRNAs) are widely in use in mammalian RNAi experiments. The present study assesses the capability of 43 and 63bp dsRNAs with two 2nt long 3'-overhangs to induce RNAi in mammalian and Drosophila cells. Human Dicer was found to cleave these dsRNAs from their ends to generate two or three monomeric siRNA units, each 21-22bp in length. When, in 43bp dsRNA, there was present a highly-effective siRNA sequence in frame with respect to the Dicer digestion, considerably high RNAi activity was noted to be induced in mouse embryonic stem E14TG2a, human HeLa, Chinese hamster CHO-K1 or Drosophila S2 cells. In contrast, RNAi depending on 63bp dsRNA, containing a highly effective siRNA sequence in frame with respect to Dicer digestion, varied considerably depending on cell lines used. While there was no appreciable RNAi in HeLa cells associated with relatively strong interferon response, a significant level of RNAi was noted in E14TG2a, CHO-K1 and S2 cells, in all of which interferon response induction was but slight, if at all. It would thus follow that siRNA oligomers with sequence of a highly functional siRNA monomer unit in frame with respect to dicer digestion should serve as a good RNAi agent in Drosophila and certain mammalian cells.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"1 2","pages":"79-87"},"PeriodicalIF":0.0,"publicationDate":"2005-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737202/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28417385","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

Design of efficient DNAzymes against muscle AChR alpha-subunit cRNA in vitro and in HEK 293 cells. 体外和HEK 293细胞中抗肌AChR α亚基cRNA的高效DNAzymes的设计。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-10-14

Amr Abdelgany, M Khabir Uddin, Matthew Wood, Kazunari Taira, David Beeson

{"title":"Design of efficient DNAzymes against muscle AChR alpha-subunit cRNA in vitro and in HEK 293 cells.","authors":"Amr Abdelgany, M Khabir Uddin, Matthew Wood, Kazunari Taira, David Beeson","doi":"","DOIUrl":"","url":null,"abstract":"DNAzymes are catalytic DNA which bind to target RNA by complementary sequence arms on a Watson-Crick basis and cleave RNA at specific sites. Potential therapeutic applications require DNAzymes that can efficiently cleave their target. Here we investigate factors affecting DNAzyme cleavage efficacy against the muscle acetylcholine receptor (AChR) alpha-subunit. The 10-23 DNAzymes cleave at Y-R nucleotide motifs, where R is A or G, and Y is U or C. Targeting a series of sites within different regions of the full-coding length cRNA under simulated physiological conditions found that the most efficient motifs for cleavage were in the hierarchy: GU >/= AU > GC >>> AC. This order is consistent with the kinetic analysis of short synthetic RNA substrates that have the same binding arms but different cleavage sites. DNAzymes with longer symmetric binding arms were more efficient than those with shorter arms, while asymmetric DNAzymes with a longer arm I were also more efficient, suggesting a dominant role for arm I in determining cleavage activity. Modification of one DNAzyme by inverted thymidine (iT) or locked nucleic acids (LNA) showed the LNA-modified DNAzyme gave efficient silencing of AChR expression in HEK 293 cells. Our data demonstrate the usefulness of screening in vitro for an efficient DNAzyme prior to cellular applications.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"1 2","pages":"88-96"},"PeriodicalIF":0.0,"publicationDate":"2005-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737201/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28417386","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

Prolonged transcriptional silencing and CpG methylation induced by siRNAs targeted to the HIV-1 promoter region. 靶向HIV-1启动子区的sirna诱导的转录沉默和CpG甲基化。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-10-11

Kazuo Suzuki, Toshiaki Shijuuku, Toshihiko Fukamachi, John Zaunders, Gilles Guillemin, David Cooper, Anthony Kelleher

{"title":"Prolonged transcriptional silencing and CpG methylation induced by siRNAs targeted to the HIV-1 promoter region.","authors":"Kazuo Suzuki, Toshiaki Shijuuku, Toshihiko Fukamachi, John Zaunders, Gilles Guillemin, David Cooper, Anthony Kelleher","doi":"","DOIUrl":"","url":null,"abstract":"In addition to the degradation of homologous RNAs through the RNA interference (RNAi) pathway, small interfering RNAs (siRNAs) can in some systems induce cytosine methylation and transcriptional silencing of homologous promoters. Targeting of HIV-1 by RNAi results in transient suppression of the virus through degradation of viral transcripts. In an effort to prolong the suppressive effect of siRNAs on productive HIV-1 infection, we targeted conserved tandem NF-kappaB binding motifs in the viral LTR. A 21-nucleotide-RNA duplex induced marked and durable (at least 30 days) suppression of productive HIV-1 infection in chronically infected Magic-5 cells. This suppression is associated with CpG methylation within the 5'LTR and marked reduction of HIV-1 transcription in nuclear run-on assays. We then assessed three additional siRNAs targeting other sites within the HIV-1 promoter region. These siRNAs suppressed HIV-1 infection to different extents and the degree of suppression correlated with the extent of de novo methylation of CpG motifs within the HIV-1 promoter region. These findings indicate that HIV-1 can be silenced by an RNA-directed mechanism that suppresses transcription and induces CpG methylation. In addition to providing evidence that this RNA-directed DNA methylation is active in mammalian cells, this is the first report of prolonged suppression of HIV-1 infection induced by siRNA.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"1 2","pages":"66-78"},"PeriodicalIF":0.0,"publicationDate":"2005-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737205/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28416438","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

RNA interference: Perspectives and caveats. RNA干扰:观点和警告。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-10-07

Jens Kurreck

引用次数: 0

Frog Prince transposon-based RNAi vectors mediate efficient gene knockdown in human cells. 基于青蛙王子转座子的RNAi载体介导人类细胞中有效的基因敲除。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-09-02

Christopher D Kaufman, Zsuzsanna Izsvák, Andrea Katzer, Zoltán Ivics

{"title":"Frog Prince transposon-based RNAi vectors mediate efficient gene knockdown in human cells.","authors":"Christopher D Kaufman, Zsuzsanna Izsvák, Andrea Katzer, Zoltán Ivics","doi":"","DOIUrl":"","url":null,"abstract":"We have developed a stable RNA interference (RNAi) delivery system that is based on the Frog Prince transposable element. This plasmid-based vector system combines the gene silencing capabilities of H1 polymerase III promoter-driven short hairpin RNAs (shRNA) with the advantages of stable and efficient genomic integration of the shRNA cassette mediated by transposition. We show that the Frog Prince-based shRNA expressing system can efficiently knock down the expression of both exogenous as well as endogenous genes in human cells. Furthermore, we use the Frog Prince-based system to study the effect of knockdown of the DNA repair factor Ku70 on transposition of the Sleeping Beauty transposon. Transposon-mediated genomic integration ensures that the shRNA expression cassette and a selectable marker gene within the transposon remain intact and physically linked. We demonstrate that a major advantage of our vector system over plasmid-based shRNA delivery is both its enhanced frequency of intact genomic integration as well as higher target suppression in transgenic human cells. Due to its simplicity and effectiveness, transposon-based RNAi is an emerging tool to facilitate analysis of gene function through the establishment of stable loss-of-function cell lines.","PeriodicalId":88272,"journal":{"name":"Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research","volume":"1 2","pages":"97-104"},"PeriodicalIF":0.0,"publicationDate":"2005-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737204/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28417387","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

Mechanisms of RNAi: mRNA cleavage fragments may indicate stalled RISC. RNAi机制:mRNA裂解片段可能提示RISC停滞。

Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research Pub Date : 2005-07-28

Torgeir Holen

引用次数: 0