Method for the in vitro synthesis of short double stranded rnas

Details Method for the in vitro synthesis of short double stranded rnas Method for the in vitro synthesis of short double stranded rnas

C - Chemistry, Metallurgy

12

N

C12N 15/11 (2006.01) A61K 31/713 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2464626

The present invention relates to the field of synthesis of short double- stranded RNAs. An in vitro transcription method using bacteriophage polymerases and target sequence-specific single-stranded DNA oligonucleotides as templates is disclosed. The present invention finds particularly advantageous use in the synthesis of short interfering RNAs (siRNAs) that have been shown to function as key intermediates in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants and RNA interference in invertebrates and vertebrate systems.

L'invention concerne le domaine de la synthèse de brins d'ARN bicaténaire courts. L'invention concerne un procédé de transcription <i>in vitro</i> faisant appel à des polymérases bactériophages et à des oligonucléotides d'ADN monocaténaire spécifique à une séquence cible, en tant que modèles. L'invention permet de faire appel de manière particulièrement avantageuse à la synthèse de brins d'ARN d'interférence courts (ARNsi), s'avérant fonctionner en tant qu'intermédiaires clés dans le déclenchement de la dégradation d'ARN spécifique à une séquence lors du silençage génique post-transcriptionnel chez des végétaux et d'interférence d'ARN dans des systèmes invertébrés et vertébrés.

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