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Systematic Study Optimizing Cas12 Biosensors for Rapid SARS-CoV-2 Detection

Bender, Alexandra Rae
http://orcid.org/0000-0001-7124-5628
http://rave.ohiolink.edu/etdc/view?acc_num=miami1652221723644359

Abstract Details

, Master of Science, Miami University, Chemistry and Biochemistry.
The COVID-19 pandemic has highlighted the importance of diagnostics in detecting and containing infectious diseases. Recently, a class of CRISPR-Cas enzymes, known as Cas12, has been discovered and is being utilized as a biosensor for detecting genetic biomarkers. Cas12 is similar to other CRISPR-Cas proteins, in that it forms an RNA-protein complex with crRNA, which then recognizes and cleaves target DNA determined by the crRNA sequence. Cas12 is unique in that upon activation, it non-specifically cleaves any nearby DNA. This non-specific cleavage property can be leveraged to create molecular biosensors by adding in a fluorescent reporter DNA substrate to the reaction. In our system, Cas12 is programmed to recognize a specific sequence of the SARS-CoV-2 genome. This class of biosensor is useful due to a faster response compared to traditional PCR-based assays. However, CRISPR-Cas biosensors suffer from off-target activity, meaning that similar sequences can activate the biosensor, resulting in false positive signal. The goal of this project is to systematically study factors that affect Cas12 activity. Specifically, we investigated how the role of crRNA concentration, crRNA length, and mutation position affect Cas12 activity. Findings from this research will aid in developing next-generation point-of-care diagnostics for detecting and containing disease outbreaks.
Kevin Yehl (Advisor)
David Tierney (Committee Member)
Andrea Kravats (Committee Member)
Neil Danielson (Committee Member)
55 p.
Biology; Chemistry; Molecular Biology; Molecular Chemistry
COVID19; CRISPR-Cas12; biosensors; bioanalytical chemistry

Recommended Citations

Citations

  • Bender, A. R. (2022). Systematic Study Optimizing Cas12 Biosensors for Rapid SARS-CoV-2 Detection [Master's thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1652221723644359

    APA Style (7th edition)

  • Bender, Alexandra. Systematic Study Optimizing Cas12 Biosensors for Rapid SARS-CoV-2 Detection. 2022. Miami University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1652221723644359.

    MLA Style (8th edition)

  • Bender, Alexandra. "Systematic Study Optimizing Cas12 Biosensors for Rapid SARS-CoV-2 Detection." Master's thesis, Miami University, 2022. http://rave.ohiolink.edu/etdc/view?acc_num=miami1652221723644359

    Chicago Manual of Style (17th edition)

miami1652221723644359
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© 2022, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.