Publisher Correction: DNA synthesis technologies to close the gene writing gap

Hoose, Alex; Vellacott, Richard; Storch, Marko; Freemont, Paul S.; Ryadnov, Maxim G.

doi:10.1038/s41570-023-00521-x

Cited by 1 publication

(2 citation statements)

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“…Enzymatic DNA synthesis stands out as a green, mild, and efficient method, differing from traditional solid-state chemical synthesis. 31,32 Enzymatic DNA synthesis relies on the activity of TdT, an enzyme that facilitates the precise addition of deoxyribonucleotide triphosphates (dNTPs) to the 3′ end of a primer. 33−37 The development of an efficient high-throughput screening assay is critical to directed evolution of TdT.…”

Section: ■ Introductionmentioning

confidence: 99%

“…In recent years, researchers have been interested in the enzymatic de novo synthesis of DNA. Enzymatic DNA synthesis stands out as a green, mild, and efficient method, differing from traditional solid-state chemical synthesis. , Enzymatic DNA synthesis relies on the activity of TdT, an enzyme that facilitates the precise addition of deoxyribonucleotide triphosphates (dNTPs) to the 3′ end of a primer. − The development of an efficient high-throughput screening assay is critical to directed evolution of TdT. In this study, we have developed a high-throughput assay utilizing ADE-OPI-MS, with a rate of 3 s per sample, to accurately identify oligonucleotides, which in turn has been employed to accelerate directed evolution of TdT.…”

Section: Introductionmentioning

confidence: 99%

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High-Throughput Screening for Oligonucleotide Detection by ADE-OPI-MS

Hu,

Zhang,

et al. 2024

Anal. Chem.

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Oligonucleotides represent a class of shorter DNA or RNA nucleic acid polymers extensively applied in the biomedical field. Despite progress in detecting and analyzing oligonucleotides, high-throughput analysis of the samples remains challenging. In this work, a high-throughput analysis method for oligonucleotide analysis was developed based on acoustic droplet ejection-open port interface-mass spectrometry (ADE-OPI-MS) technology. This approach was applied to determine the enzymatic activity of terminal deoxynucleotide transferase (TdT) for DNA synthesis, with a rate of 3 s/sample, which enhanced single-sample analysis efficiency approximately 60-fold over the previous gel analysis. After testing approximately 10,000 TdT mutants, we obtained three new variants with higher catalytic activities. Finally, by integrating these mutants, the catalytic activity of TdT was improved about 4 times compared to the starting mutant. Our results successfully established a high-throughput screening method for oligonucleotide analysis, which not only provides a foundation to engineer highly efficient TdT for ab initio synthesis of DNA but also paves the way for the potential application of oligonucleotide analysis in biomedical fields.

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Section: ■ Introductionmentioning

confidence: 99%