| Téléchargement | - Voir la version finale : Forward and robust selection of the most potent and noncellular toxic siRNAs from RNAi libraries (PDF, 3.1 Mio)
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| DOI | Trouver le DOI : https://doi.org/10.1093/nar/gkn953 |
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| Auteur | Rechercher : Li, Zhen; Rechercher : Fortin, Yves1; Rechercher : Shen, Shi-Hsiang1 |
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| Affiliation | - Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
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| Format | Texte, Article |
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| Résumé | Use of highly potent small interfering RNAs (siRNAs) can substantially reduce dose-dependent cytotoxic and off-target effects. We developed a genetic forward approach by fusing the cytosine deaminase gene with targets for the robust identification of highly potent siRNAs from RNA interference (RNAi) libraries that were directly delivered into cells via bacterial invasion. We demonstrated that two simple drug selection cycles performed conveniently in a single container predominately enriched two siRNAs targeting the MVP gene (siMVP) and one siRNA targeting the egfp gene (siEGFP) in surviving cells and these proved to be the most effective siRNAs reported. Furthermore, the potent siRNAs isolated from the surviving cells possessed noncellular toxic characteristics. Interestingly, the length of highly potent siMVPs identified could be as short as 16-mer, and increasing the length of their native sequences dramatically reduced RNAi potency. These results suggest that the current approach can robustly discover the most potent and nontoxic siRNAs in the surviving cells, and thus has great potential in facilitating RNAi applications by minimizing the dose-dependent and sequence nonspecific side effects of siRNAs. |
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| Date de publication | 2008-11-29 |
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| Maison d’édition | Oxford University Press |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Numéro NPARC | 23004911 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | 3fb6fff0-43ce-4d8b-81a9-745f21f47443 |
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| Enregistrement créé | 2019-01-21 |
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| Enregistrement modifié | 2020-05-30 |
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