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Related Experiment Video

Updated: May 17, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

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Published on: March 25, 2020

Converting One In Vitro Selected DNA Molecule Into Two Bacteria-Responsive DNAzymes by Regulation of Reaction

Xiaoqian Li1, Wei Xue1, Shusen Xiao1

  • 1Dalian POCT Laboratory, Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), School of Environmental Science and Technology, Dalian University of Technology, Dalian, China.

Angewandte Chemie (International Ed. in English)
|May 15, 2026
PubMed
Summary

A single DNAzyme sequence can detect two bacterial species, Klebsiella pneumoniae and Escherichia coli, by exploiting differences in their RNase properties. This dual-function DNAzyme shows promise for diagnosing hospital-acquired pneumonia.

Keywords:
DNAzymebacteriadeoxyribozymediagnosisin vitro selection

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Bacteria-responsive DNAzymes are crucial for pathogen detection.
  • Developing selective biosensors for multiple bacterial species remains a challenge.

Purpose of the Study:

  • To develop a single DNAzyme capable of detecting two distinct bacterial species, Klebsiella pneumoniae (KP) and Escherichia coli (EC).
  • To leverage unique enzymatic properties of bacterial RNases for differential detection.

Main Methods:

  • In vitro selection was used to isolate an RNA-cleaving fluorogenic DNAzyme, brRFD2.
  • Differential reaction conditions (pH and heat treatment) were employed to exploit distinct RNase properties of KP and EC.
  • Performance was evaluated using purified bacterial RNases and clinical samples.

Main Results:

  • The DNAzyme brRFD2 showed selective activation by KP RNase under acidic conditions (pH 3.6) and EC RNase under heat-treated conditions (pH 4.5).
  • Differential detection achieved high fold-changes in reaction rates for each target bacterium.
  • The DNAzyme demonstrated high sensitivity and specificity in diagnosing hospital-acquired pneumonia caused by KP and EC in clinical samples.

Conclusions:

  • A single DNAzyme sequence can be engineered to function as two distinct bacteria-responsive biosensors by manipulating reaction conditions.
  • This strategy offers a versatile platform for multiplexed bacterial detection and diagnostics.