CRISPR-Enabled Transition to Adsorption-Coupled Redox Transport: Toward the Use of Conventional Voltammetry in Electrochemical Biosensors
4 affiliations
- 1Materials Research Institute
- 2The Pennsylvania State University
- 3Department of Electrical Engineering
- 4Department of Biomedical Engineering
Abstract
Electrochemical aptamer-based (E-AB) biosensors and clustered regularly interspaced short palindromic repeats (CRISPR) systems have independently advanced molecular diagnostics; however, their integration is less explored. Here, we present a CRISPR-enabled electrochemical sensing platform that directly links CRISPR Cas12a-mediated cleavage of non-specific aptamers to a change of charge transfer at the electrode interface. Methylene blue (MB)-labeled aptamers are immobilized on a gold electrode, forming an electrochemically surface-confined system. Upon target-activated CRISPR cleavage, MB-labeled aptamers are released into the medium, transforming the system into a diffusion-controlled regime. This transition is directly captured through real-time electrochemical monitoring, where alternating current voltammetry reveals a progressive decrease in surface-bound redox reporters, while cyclic voltammetry simultaneously shows an increase in faradaic current due to the accumulation of freely diffusing MB species. Scan rate-dependent analysis confirms a shift from non-ideal surface-limited behavior to diffusion-controlled kinetics, consistent with the Randles–Ševčík relationship. We propose an adsorption-coupled transport model, which supports this interpretation by showing that the plausible post-cleavage voltammetric response is best described by near-surface confinement of released MB-tagged fragments rather than by pure bulk diffusion alone. Dose-response analysis shows that cyclic voltammetry achieves a improved limit of detection (LOD) of approximately , outperforming square wave voltammetry ( across a broader – range), underscoring CV as the more sensitive readout for quantitative biosensing with this platform. By leveraging the exceptional molecular specificity of CRISPR alongside the high sensitivity of E-AB sensors, this work establishes a new sensing paradigm in which conventional steady-state voltammetry can be used for analytical detection.
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BibTeX
@misc{khamsi2026crisprenabled,
title = {{CRISPR-Enabled Transition to Adsorption-Coupled Redox Transport: Toward the Use of Conventional Voltammetry in Electrochemical Biosensors}},
author = {Pouya Soltan Khamsi and Aida Ebrahimi},
howpublished = {ChemRxiv},
year = {2026},
month = sep,
doi = {10.26434/chemrxiv.15009466/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009466/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - CRISPR-Enabled Transition to Adsorption-Coupled Redox Transport: Toward the Use of Conventional Voltammetry in Electrochemical Biosensors
AU - Pouya Soltan Khamsi
AU - Aida Ebrahimi
PY - 2026
DA - 2026/09/24
PB - ChemRxiv
DO - 10.26434/chemrxiv.15009466/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009466/v1
AB - Electrochemical aptamer-based (E-AB) biosensors and clustered regularly interspaced short palindromic repeats (CRISPR) systems have independently advanced molecular diagnostics; however, their integration is less explored. Here, we present a CRISPR-enabled electrochemical sensing platform that directly links CRISPR Cas12a-mediated cleavage of non-specific aptamers to a change of charge transfer at the electrode interface. Methylene blue (MB)-labeled aptamers are immobilized on a gold electrode, forming an electrochemically surface-confined system. Upon target-activated CRISPR cleavage, MB-labeled aptamers are released into the medium, transforming the system into a diffusion-controlled regime. This transition is directly captured through real-time electrochemical monitoring, where alternating current voltammetry reveals a progressive decrease in surface-bound redox reporters, while cyclic voltammetry simultaneously shows an increase in faradaic current due to the accumulation of freely diffusing MB species. Scan rate-dependent analysis confirms a shift from non-ideal surface-limited behavior to diffusion-controlled kinetics, consistent with the Randles–Ševčík relationship. We propose an adsorption-coupled transport model, which supports this interpretation by showing that the plausible post-cleavage voltammetric response is best described by near-surface confinement of released MB-tagged fragments rather than by pure bulk diffusion alone. Dose-response analysis shows that cyclic voltammetry achieves a improved limit of detection (LOD) of approximately , outperforming square wave voltammetry ( across a broader – range), underscoring CV as the more sensitive readout for quantitative biosensing with this platform. By leveraging the exceptional molecular specificity of CRISPR alongside the high sensitivity of E-AB sensors, this work establishes a new sensing paradigm in which conventional steady-state voltammetry can be used for analytical detection.
ER -