A designed RNA selection: establishment of a stable complex between a target and selectant RNA via two coordinated interactions.

Abstract:

:In this paper, we describe a new method for selecting RNA aptamers that cooperatively bind to two specific sites within a target RNA. We designed a selection system in which two RNAs, a target RNA and a RNA pool, were assembled by employing a pre-organized GAAA tetraloop-11-nt receptor interaction. This allows us to select the binding sequence against a targeted internal loop as well as a linker region optimized for binding of the two binding sites. After the selection, the aptamers bound with dissociation constants in the nanomolar range, thereby forming a stable complex with the target RNA. Thus this method enables identification of aptamers for a specific binding site together with a linker for cooperative binding of the two RNAs. It appears that our new method can be applied generally to select RNAs that adhere tightly to a target RNA via two specific sites. The method can also be applicable for further engineering of both natural and artificial RNAs.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Shiohara T,Saito H,Inoue T

doi

10.1093/nar/gkn1012

subject

Has Abstract

pub_date

2009-02-01 00:00:00

pages

e23

issue

3

eissn

0305-1048

issn

1362-4962

pii

gkn1012

journal_volume

37

pub_type

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