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siRNA Modified with 2′‐Deoxy‐2′‐ C ‐methylpyrimidine Nucleosides
Author(s) -
Dellafiore María,
Aviñó Anna,
Alagia Adele,
Montserrat Javier M.,
Iribarren Adolfo M.,
Eritja Ramon
Publication year - 2018
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.201800077
Subject(s) - hela , chemistry , thymidine , stereochemistry , transfection , luciferase , ic50 , microbiology and biotechnology , biochemistry , dna , in vitro , biology , gene
Abstract (2′ S )‐2′‐Deoxy‐2′‐ C ‐methyluridine and (2′ R )‐2′‐deoxy‐2′‐ C ‐methyluridine were incorporated in the 3′‐overhang region of the sense and antisense strands and in positions 2 and 5 of the seed region of siRNA duplexes directed against Renilla luciferase, whereas (2′ S )‐2′‐deoxy‐2′‐ C ‐methylcytidine was incorporated in the 6‐position of the seed region of the same constructions. A dual luciferase reporter assay in transfected HeLa cells was used as a model system to measure the IC 50 values of 24 different modified duplexes. The best results were obtained by the substitution of one thymidine unit in the antisense 3′‐overhang region by (2′ S )‐ or (2′ R )‐2′‐deoxy‐2′‐ C ‐methyluridine, reducing IC 50 to half of the value observed for the natural control. The selectivity of the modified siRNA was measured, it being found that modifications in positions 5 and 6 of the seed region had a positive effect on the ON/OFF activity.