Biomics

The year 2025 marks several anniversaries directly or indirectly related to oligonucleotide synthesis. Seventy years ago, in 1955, a paper was published describing the synthesis of the dinucleotides d(TpT) and d(pTpT). In that year, a paper was published reporting the isolation of the enzyme polynucleotide phosphorylase, which polymerizes RNA in vitro, from the bacterium Azotobacter vinelandii. In the 1970s, this enzyme was used for some time to synthesize short oligonucleotides with a given sequence. The chemical synthesis of oligonucleotides has undergone significant evolution over the years, from phosphotriester, through phosphonate, phosphodiester, phosphitetriester, and then converted to phosphoramidite. The latter is currently the primary method for producing oligonucleotides using automated DNA synthesizers. In fact, the phosphoramidite method of oligonucleotide synthesis currently provides a solution to the relevant scientific and practical problems. However, the need for long-term storage of non-biological information in DNA molecules (oligonucleotides) necessitates their production in significant quantities and of a different quality (longer), which the existing approach is no longer able to handle. A solution could be provided by the enzymatic production of oligonucleotides using terminal deoxynucleotidyl transferase, an enzyme isolated from calf thymus in 1960 that enables non-template DNA synthesis.

oligonucleotide, chemical synthesis, DNA synthesizer, amidophosphite method, enzymatic synthesis, polynucleotide phosphorylase, terminal deoxynucleotidyltransferase, TdT

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