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Cellular Dynamics of Antisense Oligonucleotides and Short Interfering RNAs
Author(s) -
LEE LI KIM,
DUNHAM BRANDEE M.,
LI ZHUTING,
ROTH CHARLES M.
Publication year - 2006
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1348.061
Subject(s) - chinese hamster ovary cell , small interfering rna , oligonucleotide , transfection , rna , microbiology and biotechnology , messenger rna , green fluorescent protein , biology , rna interference , flow cytometry , chemistry , cell culture , gene , biochemistry , genetics
We aim to compare quantitatively the dynamics of the effectiveness of antisense oligonucleotides (AS ODNs) versus short interfering RNAs (siRNAs) and relate their effectiveness to sequence metrics (e.g., predicted free energy of binding). AS ODNs against a quantitative model target, pd1EGFP (destabilized enhanced GFP [green fluorescent protein]), were selected using our thermodynamic model, and siRNA sequences were designed to be identical to the AS ODN sequences in the antisense strand. We evaluated d1EGFP inhibition in transiently and stably transfected Chinese hamster ovary (CHO) cells over time using flow cytometry. Overall, our results show that the rationally designed AS ODN and siRNA sequences proved effective inhibitors of GFP expression and suggest that certain regions of mRNA may be susceptible to both AS ODNs and siRNAs.