RNA-protein hybrid ribozymes that efficiently cleave any mRNA independently of the structure of the target RNA.

Abstract:

:Ribozyme activity in vivo depends on achieving high-level expression, intracellular stability, target colocalization, and cleavage site access. At present, target site selection is problematic because of unforeseeable secondary and tertiary RNA structures that prevent cleavage. To overcome this design obstacle, we wished to engineer a ribozyme that could access any chosen site. To create this ribozyme, the constitutive transport element (CTE), an RNA motif that has the ability to interact with intracellular RNA helicases, was attached to our ribozymes so that the helicase-bound, hybrid ribozymes would be produced in cells. This modification significantly enhanced ribozyme activity in vivo, permitting cleavage of sites previously found to be inaccessible. To confer cleavage enhancement, the CTE must retain helicase-binding activity. Binding experiments demonstrated the likely involvement of RNA helicase(s). We found that attachment of the RNA motif to our tRNA ribozymes leads to cleavage in vivo at the chosen target site regardless of the local RNA secondary or tertiary structure.

authors

Warashina M,Kuwabara T,Kato Y,Sano M,Taira K

doi

10.1073/pnas.091411398

keywords:

subject

Has Abstract

pub_date

2001-05-08 00:00:00

pages

5572-7

issue

10

eissn

0027-8424

issn

1091-6490

pii

98/10/5572

journal_volume

98

pub_type

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