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. 2024 Feb 13;13(3):837–850. doi: 10.1021/acssynbio.3c00655

Figure 1.

Figure 1

CRISPRD workflow for multiplex detection of hrHPVs. (a) CRISPRD is designed to detect three targets (HPV16, HPV16, and an internal control) simultaneously. The two layers of amplification, LAMP for DNA and T7-based transcription for RNA, are followed by CRISPR-based specific detection of the three targets in one pot. CRISPRD deploys three orthogonal Cas nucleases (TccCas13a, HheCas13a, and AapCas12b) to detect their cognate target via gRNAs and digest its associated fluorophore-conjugated DNA or RNA reporter only. Target-induced Cas variant-based digestion of the reporter releases the fluorophore as a visual fluorescent readout for pathogen detection. (b) From sample to answer: after processing the sample, nucleic acid is loaded to the one-pot CRISPRD reaction where amplification and detection occur in a single step.