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Technologies for Emerging Infectious Diseases

Genetic tools

Elucidating the Role of CRISPR/Cas in Single-Step Isothermal Nucleic Acid Amplification Testing Assays

Friday, October 13, 2023
8:15 AM – 8:30 AM PDT
Location: 323 (Center, Level 3)

    Presenting Author(s)

  • Fangchi Shao, BSE

    Ph.D. Candidate
    Johns Hopkins University
    BALTIMORE, Maryland, United States

    • Co-Author(s)

  • JP

    Joon Soo Park

    Johns hopkins university, United States

  • GZ

    Guojie Zhao

    Johns Hopkins University, United States

  • KH

    Kuangwen Hsieh

    Assistant Research Professor
    Johns Hopkins University, United States

    • Primary Investigator(s)

  • JW

    Jeff Tza-Huei Wang

    Principal Investigator
    Johns Hopkins University, United States

This study present a systemic investigation of the role and impact of CRISPR/Cas in the assay performance of single-step CRISPR/Cas-assisted isothermal nucleic acid amplification assay by developing and comparing three assays with a common isothermal amplification, recombinase polymerase amplification, but different CRISPR/Cas12a methods. We found that the addition of CRISPR/Cas12a indeed unlocks signal amplification but, at the same time, impedes RPA and that CRISPR/Cas12a concentration is a key parameter for attenuating RPA impediment and ensuring assay performance.