A rapid nucleic acid detection platform based on phosphorothioate-DNA and sulfur binding domain

Nucleic acid detection plays a key role in diverse diagnosis and disease control.Currently available nucleic acid detection techniques are challenged by trade-offs among speed,simplicity,precision and cost.Here,we described a novel method,designated SENSOR(Sulfur DNA mediated nucleic acid sensing platform),for rapid nucleic acid detection.SENSOR was developed from phosphorothioate(PT)-DNA and sulfur binding domain(SBD)which specifically binds double-stranded PT-modified DNA.SENSOR utilizes PT-DNA oligo and SBD as targeting module,which is linked with split luciferase reporter to generate luminescence signal within 10 min.We tested detection on synthesized nucleic acid and COVID-19 pseudovirus,achieving attomolar sensitivity combined with an amplification procedure.Single nucleotide polymorphisms(SNP)could also be discriminated.Indicating SENSOR a new promising nucleic acid detection technique.

DNA硫修饰结合蛋白识别序列的表征与拓展促进新型核酸检测技术发展

DNA硫修饰是一种DNA磷酸骨架上的非桥接氧原子被硫原子所取代的修饰,属于原核生物表观遗传标志物之一,它对细菌防御系统、抗氧化应激和基因调控等生命过程具有重要影响.DNA硫修饰可以被硫修饰依赖限制性核酸内切酶的硫结合域SBD特异性识别,这使其具有被开发为新型生物技术工具的潜力.然而目前关于SBD蛋白识别序列的研究仍比较粗浅且仅限于少数天然序列模式,这极大限制了其应用开发.因此,我们建立了一种针对SBD蛋白识别序列表征的技术(SBS-seq),用于对SBD的完整识别序列进行高分辨率表征.而且,采用在DNA上串联多个硫修饰的方案来提高SBD的结合亲和力并拓展了其序列识别范围.在此基础上,我们进一步优化了基于SBD和硫修饰DNA的新型核酸检测技术.本工作为硫修饰依赖限制性核酸内切酶的研究提供了思路和范例,并促进了硫修饰DNA相关生物技术的应用.