Synthesis and Characterization of Chimeric 2‐5A‐DNA Oligonucleotides

Abstract

This unit provides protocols for the synthesis and characterization of 2‐5A‐antisense nucleic acids. These chimeric oligonucleotides consist of 2′,5′‐phosphodiester‐linked oligoadenylates ligated to 3′,5′‐deoxyribonucleotides and are readily prepared using phosphoramidite chemistry on CPG solid supports. The 3′,5′‐deoxyribonucleotide functions as the antisense domain to target a given mRNA sequence, while the 2′,5′‐phosphodiester‐linked oligoadenylate serves to locally activate 2‐5A‐dependent RNase L, causing the targeted sequence to be cleaved.