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Specific 3′‐Terminal Modification of DNA with a Novel Nucleoside Analogue that Allows a Covalent Linkage of a Nuclear Localization Signal and Enhancement of DNA Stability
Author(s) -
Ikeda Yutaka,
Kawahara Shunichi,
Yoshinari Koichi,
Fujita Satoshi,
Taira Kazunari
Publication year - 2005
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200400142
Subject(s) - dna , maleimide , terminal deoxynucleotidyl transferase , covalent bond , chemistry , nucleoside , rna , biochemistry , combinatorial chemistry , organic chemistry , apoptosis , tunel assay , gene
Abstract We report a straightforward method for the site‐specific modification of long double‐stranded DNA by using a maleimide adduct of deoxycytidine. This novel nucleoside analogue was efficiently incorporated at the 3′‐termini of DNA by terminal deoxynucleotidyl transferase (TdT). Thiol‐containing compounds can be covalently linked to the maleimide moieties. We added a nuclear localization signal peptide to the 3′‐terminal of a 350 bp‐long DNA that encoded short‐hairpin RNA, and these modifications resulted in the enhancement of silencing activity by RNA interference. This enhancement is mainly attributed to increased stability of the template DNA.