Premium
Efficient Synthesis of Light‐Triggered Circular Antisense Oligonucleotides Targeting Cellular Protein Expression
Author(s) -
Yang Linlin,
Kim Hyun Bum,
Sul JaiYoon,
Yeldell Sean B.,
Eberwine James H.,
Dmochowski Ivan J.
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.201800012
Subject(s) - oligonucleotide , linker , chemistry , rna , biophysics , cycloaddition , dna , glial fibrillary acidic protein , microbiology and biotechnology , biochemistry , biology , gene , immunohistochemistry , computer science , immunology , catalysis , operating system
Light‐activated (“caged”) antisense oligonucleotides are powerful molecules for regulating gene expression at submicron spatial resolution through the focal modulation of endogenous cellular processes. Cyclized caged oligos are particularly promising structures because of their inherent stability and similarity to naturally occurring circular DNA and RNA molecules. Here, we introduce an efficient route for cyclizing an antisense oligodeoxynucleotide incorporating a photocleavable linker. Oligo cyclization was achieved for several sequences in nearly quantitative yields through intramolecular copper(I)‐catalyzed azide–alkyne cycloaddition (CuAAC). Caging stability and light activation were characterized by FRET efficiency, denaturing gel assay, and melting temperature measurements. Finally, a cyclized caged oligo was designed to target gfap , and it gave a tenfold reduction in glial fibrillary acidic protein upon photoactivation in astrocytes.