Print Email Facebook Twitter Detection of CRISPR-dCas9 on DNA with Solid-State Nanopores Title Detection of CRISPR-dCas9 on DNA with Solid-State Nanopores Author Yang, W.W.W. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Restrepo Perez, L. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft) Bengtson, M.L. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Heerema, S.J. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Birnie, A.T.F. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) van der Torre, J. (TU Delft BN/Technici en Analisten; Kavli institute of nanoscience Delft) Dekker, C. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Date 2018 Abstract Solid-state nanopores have emerged as promising platforms for biosensing including diagnostics for disease detection. Here we show nanopore experiments that detect CRISPR-dCas9, a sequence-specific RNA-guided protein system that specifically binds to a target DNA sequence. While CRISPR-Cas9 is acclaimed for its gene editing potential, the CRISPR-dCas9 variant employed here does not cut DNA but instead remains tightly bound at a user-defined binding site, thus providing an excellent target for biosensing. In our nanopore experiments, we observe the CRISPR-dCas9 proteins as local spikes that appear on top of the ionic current blockade signal of DNA molecules that translocate through the nanopore. The proteins exhibit a pronounced blockade signal that allows for facile identification of the targeted sequence. Even at the high salt conditions (1 M LiCl) required for nanopore experiments, dCas9 proteins are found to remain stably bound. The binding position of the target sequence can be read from the spike position along the DNA signal. We anticipate applications of this nanopore-based CRISPR-dCas9 biosensing approach in DNA-typing based diagnostics such as quick disease-strain identification, antibiotic-resistance detection, and genome typing. Subject biosensingCRISPR-Cas9diagnosticsNanopores To reference this document use: http://resolver.tudelft.nl/uuid:d0d99406-5b8f-4292-89c0-cf02de707903 DOI https://doi.org/10.1021/acs.nanolett.8b02968 ISSN 1530-6984 Source Nano Letters: a journal dedicated to nanoscience and nanotechnology Part of collection Institutional Repository Document type journal article Rights © 2018 W.W.W. Yang, L. Restrepo Perez, M.L. Bengtson, S.J. Heerema, A.T.F. Birnie, J. van der Torre, C. Dekker Files PDF acs.nanolett.8b02968.pdf 1.64 MB Close viewer /islandora/object/uuid:d0d99406-5b8f-4292-89c0-cf02de707903/datastream/OBJ/view