Fast-responding functional DNA superstructures for stimuli-triggered protein release.

Yuxin Zhang,Yingfu Li,Meng Liu,Fang Cheng,Qiang Zhang,Yangyang Chang

doi:10.1039/d1sc00795e

Yuxin Zhang, Yingfu Li + Show 4 more

Open Access

https://doi.org/10.1039/d1sc00795e

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Abstract

Strategies that speed up the on-command release of proteins (e.g., enzymes) from stimuli-responsive materials are intrinsically necessary for biosensing applications, such as point-of-care testing, as they will achieve fast readouts with catalytic signal-amplification. However, current systems are challenging to work with because they usually exhibit response times on the order of hours up to days. Herein, we report on the first effort to construct a fast-responding gating system using protein-encapsulating functional DNA superstructures (denoted as protein@3D DNA). Proteins were directly embedded into 3D DNA during the one-pot rolling circle amplification process. We found that the specific DNA–DNA interaction and aptamer–ligand interaction could act as general protocols to release the loaded proteins from 3D DNA. The resulting gating system exhibits fast release kinetics on the order of minutes. Taking advantage of this finding, we designed a simple paper device by employing protein@3D DNA for colorimetric detection of toxin B (Clostridium difficile marker). This device is capable of detecting 0.1 nM toxin B within 16 minutes.

Highlights

Learning from Mother Nature has inspired the design of smart, intelligent or stimuli-responsive materials.[1]
We report on the first effort to construct a fast-responding gating system using protein-encapsulating functional DNA superstructures
Protein@3D DNA is synthesized in two steps: (1) the use of f29 DNA polymerase (Polf29), a circular template (Ct) and a DNA primer (Dp) to encapsulate proteins into 3D DNA via one-pot rolling circle ampli cation (RCA, Fig. S1†);[9] (2) isolation of protein@3D DNA by a centrifugal separation method

Summary

Introduction

Learning from Mother Nature has inspired the design of smart, intelligent or stimuli-responsive materials.[1]. The simple physical mixtures of 3D DNA with HRP resulted in the encapsulation of HRP into 3D DNA, producing a signi cantly decreased loading capacity of (0.3 Æ 0.1) Â 104 F-HRP molecules per particle.

Results

Conclusion