Multiplexed CRISPR/Cas9 quantifications based on bioinspired photonic barcodes

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated nucleases (CRISPR/Cas) platforms have a demonstrated value in nucleic acid detection, while their multiplex screening capability is still challenging. Herein, we present a novel CRISPR/Cas9 platform based on bioinspired photonic crystal (PhC) barcodes for multiplex and sensitive nucleic acid detection. The bioinspired PhC barcodes not only exhibit distinctive structural color as encoding elements, but also possess abundant functional surface groups for probe immobilization with the presence of polydopamine (PDA) coating. The CRISPR/Cas9 system recognizes and cleaves target DNA and with the help of Klenow fragment to produce ssDNA, which was subsequently captured by the PhC barcodes and the detection signal was amplified by hybridization chain reaction (HCR). Based on this platform, we demonstrated that Human Papilloma Virus (HPV) nucleic acids could be detected with a highly sensitive limit of 0.025 pM and be multiplexed assayed with high accuracy and specificity. Thus, we believe that this platform paves an avenue for multiplexed biomarkers quantification in clinical disease diagnostics. (c) 2021 Elsevier Ltd. All rights reserved.