文献类型：期刊文献

英文题名：Machine learning-optimized long single-stranded DNA synthesis technology empowers high-precision diagnostic-therapeutic integration in living cells

作者：Zhang, Yue[1,2];Xu, Yiming[1,2];Ding, Zhihua[2];Cheng, Yutian[1,2];Gao, Yu[1,2];Ye, Jiang[1,2];Zhang, Huizhan[1,2];Li, Pengfei[1,3];Wu, Haizhen[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Dept Appl Biol, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Fermentat Engn Expt Teaching Demonstrat Ctr, Shanghai 200237, Peoples R China

年份：2026

卷号：54

期号：4

外文期刊名：NUCLEIC ACIDS RESEARCH

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001687159000001)】；

基金：This work was supported by the National Key Research and Development Program of China (2021YFC2100600) and National Natural Science Foundation of China (31872608). Funding to pay the Open Access publication charges for this article was provided by National Natural Science Foundation of China (31872608).

语种：英文

摘要：While DNA nanotechnology holds transformative potential across biomedical and information storage applications, current technologies face critical limitations in synthesizing long single-stranded DNA (ssDNA) with high purity and homogeneity. To address these challenges, we developed Ouroborosyn-ssDNA, a nicking enzymatic assisted replication (NEAR) platform that synergizes enzymatic engineering with computational optimization. By integrating phi29 DNA polymerase and Nb.BbvCI nickase in formate-based buffers, we achieved extended ssDNA synthesis up to 15 000 nt while preserving sequence fidelity, resulting in a 4.73-fold yield enhancement compared to commercial buffers. Notably, machine learning-guided parameter optimization identified magnesium ion dynamics and thermal modulation as pivotal determinants of enzymatic efficiency. Furthermore, solid-phase synthesis using thiol-gold immobilized templates demonstrated 86.38% purification recovery via automated magnetic bead systems, enabling scalable production. To validate functional utility, we engineered six-helix bundle DNA origami-CRISPR complexes that achieved nucleolin-targeted genome editing in cervical cancer cells, coupling GFP-based diagnostics with therapeutic E7 oncogene disruption. These advancements directly overcome key limitations in enzymatic stochasticity and product heterogeneity through buffer engineering and computational optimization, establishing a scalable pathway for applications in precision nanomedicine, synthetic biology, and molecular data storage. This integrated strategy advances DNA nanotechnology from proof-of-concept studies toward standardized biomanufacturing of sequence-defined macromolecular architectures.