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Methods Mol Biol. 2010;623:67-79. doi: 10.1007/978-1-60761-588-0_4.

Allele-specific silencing by RNA interference.

Author information

  • National Institute of Neuroscience, NCNP, Tokyo, Japan.

Abstract

Allele-specific gene silencing by RNA interference (RNAi) is therapeutically useful for specifically inhibiting the expression of disease-associated alleles without suppressing the expression of corresponding wild-type alleles. To realize such allele-specific RNAi (ASP-RNAi), the design and assessment of small interfering RNA (siRNA) duplexes conferring ASP-RNAi is vital; however, it is also difficult. We have developed an assay system with reporter alleles that encode the Photinus and Renilla luciferase genes carrying mutant and wild-type allelic sequences in their 3'-untranslated regions. The assay system allows us to evaluate designed siRNAs and also short hairpin RNAs for allele-specific silencing against target mutant alleles as well as off-target silencing against corresponding wild-type alleles simultaneously in a qualitative and quantitative manner.

PMID:
20217544
[PubMed - indexed for MEDLINE]
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