Constructing an efficient trans-acting genomic HDV ribozyme.

Abstract:

:We have engineered a genomic HDV ribozyme to construct several trans-acting ribozymes for use in trans to cleave target RNAs. Among the 10 different combinations attempted, only HDV88-Trans had cleavage activity on the 13-nucleotide substrate, R13, in vitro. To improve the cleavage efficiency, at least in vitro, of the HDV88-Trans ribozyme (kclv = 0.022 min(-1)), we have constructed several variants that differ in forming stem II (length) in the pseudoknot secondary structure model. When cleavage rate constants were analyzed and compared among variants of HDV88-Trans, HDV88-Trans-4 yielded kclv = 1.7 min(-1). HDV88-Trans-4 thus represents the highest active genomic HDV ribozyme that functions in trans thus far constructed, and has activity under physiological conditions (pH 7.1 at 37 degrees C with 1 mM of MgCl2).

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Kawakami J,Yuda K,Suh YA,Kumar PK,Nishikawa F,Maeda H,Taira K,Ohtsuka E,Nishikawa S

doi

10.1016/0014-5793(96)00941-6

subject

Has Abstract

pub_date

1996-09-30 00:00:00

pages

132-6

issue

2

eissn

0014-5793

issn

1873-3468

pii

0014-5793(96)00941-6

journal_volume

394

pub_type

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