Ex-situ DNA synthesis accelerated

Jayateerth S Bhavikatti, CSIR-National Chemical Laboratory, Pune.

We have recently come across that the Nobel Prize for Chemistry 2020 is awarded to Emmanuelle Charpentier and Jennifer A. Doudna for developing CRISPR/Cas9 genetic scissors. This process requires guide RNA for cutting DNA at specific locations in the genome. Many techniques in molecular biology require template nucleotide strand(s). So far, most people use a chemical technique namely phosphoramidite synthesis for synthesizing defined nucleotide sequences and this method has an efficiency of 99.2-99.3%. This technique does not aid in synthesizing longer sequences although (beyond 120 bases). Also, this method involves chemicals that are considered hazardous by the US Environmental Protection Agency. 

Biology is full of wonders and living systems are marvellous. A poorly studied enzyme, terminal deoxynucleotidyl transferase (TdT), is now serving the purpose of controlled nucleotide synthesis. It is reported to expand the diversity of epitopes recognized by the B and T cells responsible for immunity. Three companies – Molecular Assemblies, DNA Script, and Nuclera are producing enzymatically synthesized DNA (using TdT) to sizes larger than 280-base sequences with 99.7-99.9% efficiency! “The synthesis has now entered the center stage,” says J. W. Efacavitch from Molecular Assemblies. The current challenge is to produce more gene- and genome-sized sequences to aid research and also use DNA for data storage.

Also read: Can bacterial toxins show a positive or healing effect?

Reference: Michael Eisenstein. Enzymatic DNA synthesis enters new phase. (2020) Nature Biotechnology 38:1113–1115. https://doi.org/10.1038/s41587-020-0695-9

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