Protocol for high-precision CRISPR-Cas12a-based SNV detection on synthetic DNA, cell line cfDNA models, and liquid biopsies
CRISPR-based diagnostics (CRISPRdx) offer promising tools for rapid and cost-effective genetic testing, but achieving single-nucleotide fidelity remains a challenge. Here, we present a protocol for high-precision detection of single-nucleotide variants (SNVs) using a Cas12a-based approach. We describe how to apply our publicly available ARTEMIS algorithm to identify targetable SNVs, design optimized CRISPR RNAs (crRNAs), and perform fluorescence-based CRISPRdx assays on synthetic DNA, cell line-derived cell-free DNA (cfDNA), and liquid biopsy samples. For complete details on the use and execution of this protocol, please refer to Kohabir et al.<sup>1</sup>.
Authors
- Jasper Linthorst (ORCID: https://orcid.org/0000-0002-8067-1087)
- Rob M.F. Wolthuis (ORCID: https://orcid.org/0000-0002-3109-1588)
- Kavish A.V. Kohabir (ORCID: https://orcid.org/0000-0002-1124-0670)
- Erik A. Sistermans (ORCID: https://orcid.org/0000-0001-7187-4563)
Institutions
- Amsterdam Neuroscience (NL)
- Amsterdam University Medical Centers (NL)
- Vrije Universiteit Amsterdam (NL)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-03-14
- DOI
- https://doi.org/10.1016/j.xpro.2025.103696
- Citations
- 1
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- University of Rochester
- Amsterdam University Medical Centers