Electronically controlled deprotection chemistry for multiplex enzymatic DNA synthesis on a chip with single-base resolution.

Authors

Lihuan Zhao, Qinzhuo Sun, Jian-Qiao Jiang, Xuezheng Wu, Yiming Dong, Dan Wu, Lin-Sheng Wu, Xin Zhao

Published in

Lab on a chip. Sep 05, 2025. Epub Sep 05, 2025.

Abstract

Enzymatic deoxyribonucleic acid (DNA) synthesis (EDS) is an environmentally friendly approach capable of generating longer and more complex sequences than chemical synthesis, making it a promising next-generation technology for high-throughput single-stranded DNA production. However, precise sequence control at high throughput remains a key challenge. Here, we present a novel electronically controlled deprotection chemistry (ECDC) integrated with a hydrogel-primer modification system on-chip for efficient multiplexed EDS. Electrochemically generated HNO₂ at the working electrodes selectively converts the 3'-oxyamino group of DNA into a hydroxyl group, enabling precise spatiotemporal control of a multipixel synthesis array and facilitating future automation. This platform enables parallel EDS with single-base resolution on silicon chips. In four-nucleotide validation experiments, single-sequence synthesis could achieve 100% accuracy, while dual-sequence synthesis reached an average accuracy of approximately 96%. Our approach provides a highly accurate solution for high-throughput ssDNA synthesis, laying the foundation for scalable and automated enzymatic DNA manufacturing.

PMID:
40911197
Bibliographic data and abstract were imported from PubMed on 05 Sep 2025.

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