Cellular DNA and RNA are made up of ribo- and 2'-deoxyribonucleic acids linked together via 3'-5' phosphodiester linkages and by far comprise the bulk of polynucleic acids found in cells. Much less common are oligonucleotides which have 2'-5' linkages. However, a unique feature of 2'-5' linked oligonucleotides is their ability to bind selectively to complementary RNA. These features suggest a number of interesting uses for 2'-5' linked oligos such as their use as RNA specific probes or in antisense oligos. Recently, oligos have been synthesized using 3'-deoxy-2'-phosphoramidites and 2'-deoxy-3'-phosphoramidites to produce chimeras with 2'-5' linked ends and 3'-5' linked central regions. It was found that 2'-5' phosphorothioate oligos: 1) bind selectively to complementary RNA with the same affinity as phosphodiester oligos; 2) exhibit much less nonspecific binding to cellular proteins; 3) do not activate RNase H. A 3'-deoxynucleoside at the 3'-terminus of an otherwise normal oligonucleotide effectively blocks polymerase extension.
Coupling: No changes needed from standard method recommended by synthesizer manufacturer.
Deprotection: No changes needed from standard method recommended by synthesizer manufacturer. Note: Deprotection in AMA (50:50 Ammonium hydroxide/MethylAmine) may result in about 5% N4-methyl-dC side product at the modification site.