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.
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.
Ikeda
Yutaka
Y
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, 7-3-1 Hongo, Tokyo 113-8656, Japan.
Kawahara
Shun-ichi
S
Yoshinari
Koichi
K
Fujita
Satoshi
S
Taira
Kazunari
K
D000595:Amino Acid Sequence; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000138:chemical synthesis*; D003841:Deoxycytidine / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008301:Maleimides / Q000737:chemistry*; D015394:Molecular Structure; D019913:Nuclear Localization Signals; D034622:RNA Interference
D018736:DNA Adducts; D008301:Maleimides; D019913:Nuclear Localization Signals; D003841:Deoxycytidine; D004247:DNA