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Synthesis, Gene Silencing, and Molecular Modeling Studies of 4 '-C-Aminomethyl-2 '-O-methyl Modified Small Interfering RNAs

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Title Synthesis, Gene Silencing, and Molecular Modeling Studies of 4 '-C-Aminomethyl-2 '-O-methyl Modified Small Interfering RNAs
 
Creator GORE, KR
NAWALE, GN
HARIKRISHNA, S
CHITTOOR, VG
PANDEY, SK
HOBARTNER, C
PATANKAR, S
PRADEEPKUMAR, PI
 
Subject LOCKED NUCLEIC-ACIDS
DOUBLE-STRANDED-RNA
CHEMICAL-MODIFICATION
EXONUCLEASE RESISTANCE
BIOLOGICAL-ACTIVITY
THERMAL-STABILITY
MAMMALIAN-CELLS
CELLULAR UPTAKE
FORCE-FIELD
SIRNA
 
Description The linear syntheses of 4'-C-aminomethyl-2'-O-methyl uridine and cytidine nucleoside phosphoramidites were achieved using glucose as the starting material. The modified RNA building blocks were incorporated into small interfering RNAs (siRNAs) by employing solid phase RNA synthesis. Thermal melting studies showed that the modified siRNA duplexes exhibited slightly lower T-m (similar to 1 degrees C/modification) compared to the unmodified duplex. Molecular dynamics simulations revealed that the 4'-C-aminomethyl-2'-O-methyl modified nucleotides adopt South-type conformation in a siRNA duplex, thereby altering the stacking and hydrogen-bonding interactions. These modified siRNAs were also evaluated for their gene silencing efficiency in HeLa cells using a luciferase-based reporter assay. The results indicate that the modifications are well tolerated in various positions of the passenger strand and at the 3' end of the guide strand but are less tolerated in the seed region of the guide strand. The modified siRNAs exhibited prolonged stability in human serum compared to unmodified siRNA. This work has implications for the use of 4'-C-aminomethyl-2'-O-methyl modified nucleotides to overcome some of the challenges associated with the therapeutic utilities of siRNAs.
 
Publisher AMER CHEMICAL SOC
 
Date 2014-10-16T13:25:28Z
2014-10-16T13:25:28Z
2012
 
Type Article
 
Identifier JOURNAL OF ORGANIC CHEMISTRY, 77(7)3233-3245
http://dx.doi.org/10.1021/jo202666m
http://dspace.library.iitb.ac.in/jspui/handle/100/15660
 
Language en