1. RRI Digital Repository
  2. 06. Soft Condensed Matter
  3. Research Papers (SCM)
Please use this identifier to cite or link to this item: http://hdl.handle.net/2289/8667
Full metadata record
DC FieldValueLanguage
dc.contributor.authorNagpure, Punitkumar-
dc.contributor.authorSuresh, Sarangi-
dc.contributor.authorShet, Divya-
dc.contributor.authorSoni, Gautam V-
dc.date.accessioned2026-02-12T10:24:09Z-
dc.date.available2026-02-12T10:24:09Z-
dc.date.issued2025-11-21-
dc.identifier.citationJournal of Nanobitechnology, 2025, Vol. 23 (1), AR No. 728en_US
dc.identifier.urihttp://hdl.handle.net/2289/8667-
dc.descriptionOpen Accessen_US
dc.description.abstractNanopore sensing, a high-resolution DNA sequencing technology, is rapidly expanding into novel and exciting directions of probing specific DNA-enzyme interactions. Although proven excellent for the detecting structural features of bare DNA, quantitative measurements on enzyme-DNA complexes and their real-time activity are lagging and only starting to emerge for long DNA templates. Signal-to-noise requirement and high translocation speeds make it difficult to detect protein bound on biologically relevant plasmid-length DNA. To this end we report accurate position detection of a catalytically active Cas9 bound to its single or multiple target sites on the DNA. Protein position is fingerprinted using event charge deficit (ECD) based analysis of the high signal-to-noise electrical signals as the complex translocates through a glass nanopore. Using a time-dependent assay, we quantify the kinetics of the released products upon enzymatic cleavage of the target DNA by the wild-type Cas9 nuclease. Our approach enables the nanopore-based single-molecule sensing of DNA-protein complexes, for real-time monitoring of biochemical reactions. This may help understand protein binding & localization as well as improve Cas9-based targeting in genome engineering applications.en_US
dc.language.isoenen_US
dc.publisherSpringer Natureen_US
dc.relation.urihttps://www.biorxiv.org/content/10.1101/2025.05.30.656938v1en_US
dc.relation.urihttps://doi.org/10.1186/s12951-025-03837-6en_US
dc.rights2025 The Author(s)en_US
dc.subjectDNA Nanomachinesen_US
dc.subjectDNA sequencingen_US
dc.subjectNanosensorsen_US
dc.subjectNanoporesen_US
dc.subjectProtein sequencingen_US
dc.subjectNanofabricationen_US
dc.subjectNanopatterningen_US
dc.titleNanopore assay for fingerprinting DNA binding and quantifying real-time cleavage by catalytically active Cas9 enzymeen_US
dc.typeArticleen_US
Appears in Collections:Research Papers (SCM)

Files in This Item:
File Description SizeFormat 
2025_Journal of Nanobitechnology_Vol. 23 (1)_AR No. 728Open Access2.26 MBAdobe PDFView/Open
Show simple item record


Items in RRI Digital Repository are protected by copyright, with all rights reserved, unless otherwise indicated.