Gene synthesis is the main content of current synthetic biology. Through gene synthesis, genes that do not exist in nature can be obtained, which provide a whole new direction for humans to transform organisms. In the foreseeable future, gene synthesis will play an important role in the field of life sciences. However, the current speed of gene synthesis still cannot fully meet the needs of biological development. A few days ago, a new document published by “Nature Biotechnology” pointed out that a new gene synthesis technology actually shortened the synthesis time of new genes to only one day!
Traditional gene synthesis is mostly used for the creation of antibody genes, primarily using terminal deoxynucleotide transferase (TdT), which can add new nucleotides to the oligonucleotide chain without following the DNA template chain. Thereby, new variants of millions of antibody genes have been written, and the immune system can use these variants to target invaders. But, natural TdT can only randomly add new deoxynucleotides, and researchers currently cannot accurately control the synthesis process.
The researchers of this article designed the TdT-dNTP conjugate, which can be actively covalently linked to the 3’end when incorporated into the primer. This prevents the binding of other TdT-dNTP conjugates to the primer. During further extension, the linker can be cleaved, the 3’end of the primer is deprotected, and a single nucleotide is added to the extension. This makes the “termination” process reversible from now on, and researchers can turn on or end the role of TdT as required.
The researchers have shown that the active TdT-dNTP conjugate can connect the nucleoside triphosphate to various points on the surface of the polymerase through two different linkers. They found that the TdT-dNTP conjugate can achieve DNA in 10-20s within one single nucleotide extension of the molecule. So, this cycle of extension and deprotection reaction can quickly and accurately write a defined sequence, and it takes only one day to synthesize a complete gene.
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Palluk S, Arlow DH, de Rond T, et al. De novo DNA synthesis using polymerase-nucleotide conjugates. Nature Biotechnology. 2018 Aug;36(7):645-650. DOI: 10.1038/nbt.4173. PMID: 29912208.