Synthetic Pre-miRNA-Based shRNA as Potent RNAi Triggers

Autor: Kazuya Terasawa, Kazuharu Shimizu, Gozoh Tsujimoto
Jazyk: angličtina
Rok vydání: 2011
Předmět:
Zdroj: Journal of Nucleic Acids, Vol 2011 (2011)
Druh dokumentu: article
ISSN: 2090-021X
DOI: 10.4061/2011/131579
Popis: RNA interference (RNAi) is a powerful tool for studying gene function owing to the ease with which it can selectively silence genes of interest, and it has also attracted attention because of its potential for therapeutic applications. Chemically synthesized small interfering RNAs (siRNAs) and DNA vector-based short hairpin RNAs (shRNAs) are now widely used as RNAi triggers. In contrast to expressed shRNAs, the use of synthetic shRNAs is limited. Here we designed shRNAs modeled on a precursor microRNA (pre-miRNA) and evaluated their biological activity. We demonstrated that chemically synthetic pre-miRNA-based shRNAs have more potent RNAi activity than their corresponding siRNAs and found that their antisense strands are more efficiently incorporated into the RNA-induced silencing complex. Although greater off-target effects and interferon responses were induced by shRNAs than by their corresponding siRNAs, these effects could be overcome by simply using a lower concentration or by optimizing and chemically modifying shRNAs similar to synthetic siRNAs. These are challenges for the future.
Databáze: Directory of Open Access Journals