In situ terminus-regulated DNA hydrogelation for ultrasensitive on-chip microRNA assay

Shengyuan Deng; Juan Yan; Fei Wang; Yan Su; Xueli Zhang; Qian Li; Guang Liu; Chunhai Fan; Hao Pei; Ying Wan

doi:10.1016/j.bios.2019.04.053

In situ terminus-regulated DNA hydrogelation for ultrasensitive on-chip microRNA assay

Biosens Bioelectron. 2019 Jul 15:137:263-270. doi: 10.1016/j.bios.2019.04.053. Epub 2019 May 3.

Authors

Shengyuan Deng¹, Juan Yan², Fei Wang³, Yan Su⁴, Xueli Zhang⁵, Qian Li⁶, Guang Liu¹, Chunhai Fan⁶, Hao Pei⁷, Ying Wan⁸

Affiliations

¹ School of Environmental and Biological and Engineering, Nanjing University of Science and Technology, Nanjing, 210094, China.
² College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, China.
³ Joint Research Centre for Precision Medicine, Shanghai Jiao Tong University & Affiliated Sixth People's Hospital South Campus, Fengxian Hospital, Shanghai, 201499, China; School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
⁴ School of Mechanical Engineering, Nanjing University of Science and Engineering, Nanjing, 210094, China.
⁵ Joint Research Centre for Precision Medicine, Shanghai Jiao Tong University & Affiliated Sixth People's Hospital South Campus, Fengxian Hospital, Shanghai, 201499, China.
⁶ School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
⁷ Shanghai Key Laboratory of Green Chemistry and Chemical Processes, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai, 200241, China. Electronic address: peihao@chem.ecnu.edu.cn.
⁸ School of Mechanical Engineering, Nanjing University of Science and Engineering, Nanjing, 210094, China. Electronic address: wanying@njust.edu.cn.

PMID: 31121463
DOI: 10.1016/j.bios.2019.04.053

Abstract

In this work, a dynamic terminus-regulated fabric of DNA hydrogel was invented in debt to the reiterative catalysis of terminal deoxynucleotidyl transferase (TdT). It extended free 3'-OH end to an overhang of homopolymeric adenosine base pair, and alternated with branching from the frayed complementary seed oligo T₂₀G₅. The cycle of this template-independent and isothermal amplification resulted in a microscale dendritic DNA fractal at first, which then gelatinized into a cohesive and intricate 3D network. Details of the complex were elucidated with gel electrophoresis, confocal and atomic force microscopy. Its well hydrated inner space could further provide plenty of biocompatible chambers for enzymatic transducers fused along the elongation. Taking merits of this neat and flexible setup, an in situ hydrogelation strategy was developed and utilized in the signal cascade of a miRNA biomarker detector on an electrode microarray, thus accomplished an ultrasensitive, selective and high-throughput sensing even for real samples. This collective manipulation of DNA-protein hydrogel ensemble on interface demonstrates its potency as a general scheme of sensitization in bioanalytical applications.

Keywords: DNA hydrogel; Electrochemical miRNA assay; Microarray sensor; Non-template isothermal amplification; Terminal deoxynucleotidyl transferase.

MeSH terms

Biosensing Techniques*
DNA / chemistry*
Electrochemical Techniques*
Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry
Lab-On-A-Chip Devices
Metal Nanoparticles / chemistry
MicroRNAs / chemistry
MicroRNAs / isolation & purification*

Substances

MicroRNAs
Hydrogel, Polyethylene Glycol Dimethacrylate
DNA