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| Vasques,Luciana dos Reis; Pujiz,Regiane Simoni; Strauss,Bryan Eric; Krieger,José Eduardo. |
| E2F1 plays a key role in cell-cycle regulation in mammals, since its transcription factor activity controls genes required for DNA synthesis and apoptosis. E2F1 deregulation is a common feature among different tumor types and can be a major cause of cell proliferation. Thus, blocking E2F1 expression by RNA interference represents a promising therapeutic approach. In this study, the introduction of specific short hairpin RNAs (shRNAs) reduced E2f1 expression by up to 77%, and impaired rat glioma cell proliferation by approximately 70%, as compared to control cells. Furthermore, we investigated the expression of E2f1 target genes, Cyclin A and Cyclin E. Cyclin A was found to be down-regulated, whereas Cyclin E had similar expression to control cells,... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: RNAi; ShRNA; E2F1; Proliferation; Cancer. |
| Ano: 2010 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000100005 |
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| Stove,Veronique; Smits,Kaatje; Naessens,Evelien; Plum,Jean; Verhasselt,Bruno. |
| RNA interference (RNAi), mediated by short double-stranded RNAs, is a powerful mechanism for posttranscriptional gene silencing. Sustained expression of short hairpin RNA (shRNA) can be accomplished in mammalian cells by viral delivery systems. Using lentiviral constructs, stable gene silencing is established both in dividing and non-dividing cells. Targeting one single gene can lead to the development of escape mutants or may be insufficient to silence redundant pathways. Therefore, simultaneous targeting of multiple genes may be necessary. We have generated a lentiviral vector-based system for expression of multiple shRNAs from a single viral vector, which also encodes an EGFP reporter protein. We show that knock-down of each single gene from multiple... |
| Tipo: Journal article |
Palavras-chave: EGFP; Lentiviral gene transfer; Multiple knock-down; RNAi; Rho GTPases; ShRNA. |
| Ano: 2006 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500013 |
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| Ni,Wei; Hu,Shengwei; Hazi,Wureli; Wang,Yuanzhi; Qiao,Jun; Yan,Ren; Chen,Chuangfu. |
| Background: Gene silencing mediated by small interfering RNA (siRNA) has become a powerful biological tool for the regulation of gene expression. For the synthesis of siRNA by vector-based expression systems, several mammalian small nuclear RNA (snRNA) promoters have been cloned and shown different transcriptional efficiencies. Results: In this study, we identified a sheep 7SK snRNA (s7SK) promoter based on the highly conserved polymerase III promoter elements. Promoter activity was measured by promoter-driven shRNA expression to suppress expression of an exogenous reporter gene and endogenous sheep gene. Conclusions: The knock down assay demonstrated that the s7SK induced more stronger inhibition effect than human U6 and H1 promoters. The use of this... |
| Tipo: Journal article |
Palavras-chave: 7SK promoter; RNAi; Sheep; ShRNA. |
| Ano: 2012 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000200010 |
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| Carli,Gabriel José de; Rotela,Abdon Troche; Lubini,Greice; Contiliani,Danyel Fernandes; Candia,Nidia Benítez; Depintor,Thiago S.; Abreu,Fabiano Carlos Pinto de; Simões,Zilá Luz Paulino; Ríos,Danilo Fernández; Pereira,Tiago Campos. |
| Abstract RNA interference (RNAi) is a powerful gene silencing technology, widely used in analyses of reverse genetics, development of therapeutic strategies and generation of biotechnological products. Here we present a free software tool for the rational design of RNAi effectors, named siRNA and shRNA designer (SSD). SSD incorporates our previously developed software Strand Analysis to construct template DNAs amenable for the large scale production of mono-, bi- and trivalent multimeric shRNAs, via in vitro rolling circle transcription. We tested SSD by creating a trivalent multimeric shRNA against the vitellogenin gene of Apis mellifera. RT-qPCR analysis revealed that our molecule promoted a decrease in more than 50% of the target mRNA, in a... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Gene silencing; SiRNA; Multimeric; ShRNA; Free software. |
| Ano: 2020 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572020000100802 |
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| Zhang,Shumin; Xiong,Kai; Xie,Zhourui; Nan,Wenting; Liu,Honglin; Chen,Jie. |
| β-lactoglobulin (BLG), a dominant allergen in goat milk, is difficult to remove by traditional biochemical methods. Its elimination from goat milk by genetic modification therefore poses a major challenge for modern goat breeders. A shRNA targeting BLG mRNA with high interference efficiency was identified, with which lentiviral vectors were used for mediating stable shRNA interference in goat-fetal fibroblast cells. Apart from high efficiency in the knockdown of BLG expression in these cells, lentivector-mediated RNAi manifested stable integration into the goat genome itself. Consequently, an in vitro model for goat BLG-content control was compiled, and a goat-cell line for accompanying transgenetic goat production created. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: RNAi; ShRNA; Lentivirus; BLG; Goat. |
| Ano: 2012 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000400020 |
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