University of Illinois Urbana-Champaign

CRISPR-based approaches for the detection of antimicrobial-resistant genes

Shin, Jiyong

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https://hdl.handle.net/2142/122188

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Title
CRISPR-based approaches for the detection of antimicrobial-resistant genes
Author(s)
Shin, Jiyong
Issue Date
2023-05-01
Director of Research (if dissertation) or Advisor (if thesis)
Wang, Yi-Cheng
Committee Member(s)
  • Jin, Yong-Su
  • Miller, Michael
Department of Study
Food Science & Human Nutrition
Discipline
Food Science & Human Nutrition
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
M.S.
Degree Level
Thesis
Keyword(s)
  • CRISPR biosensor
  • Cas12a
  • Nucleic acid detection
  • food safety
  • multi-drug-resistant
  • fluorescent assay
Abstract
The food industry has been seeking more advanced detection methods, and the emerging technology of biosensing based on clustered regularly interspaced short palindromic repeats (CRISPR) may hold the key to addressing this need. Here, we evaluate the current state of CRISPR technology in the realm of food safety, including the various applications and targets of each Cas effector. Although still in its early stages, CRISPR/Cas-based sensors have shown great promise and could become the new standard in food safety. Additionally, this technology has potential for use in other areas of the food industry, such as detecting antimicrobial resistant (AMR) bacteria. To this end, we developed a novel PCR-coupled CRISPR/Cas12a-based fluorescence assay capable of detecting bacteria that produce New Delhi metallo-β-lactamase (NDM). The CRISPR/Cas12a system, thanks to its specifically designed gRNA, demonstrated remarkable specificity at the single-base pair level, specifically targeting the NDM-producing genes in bacteria. Our assay demonstrated superior performance compared to previously developed real-time PCR assays and addressed important weaknesses of conventional amplification-based detection methods, such as false positives and SNP discrimination. With further research, CRISPR/Cas-based sensors have the potential to become the new gold standard for detecting AMR genes, ultimately contributing to mitigating the global threat of AMR.
Graduation Semester
2023-05
Type of Resource
Thesis
Copyright and License Information
Copyright 2023 by Jiyong Shin

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