Exploring the Trans‐Cleavage Activity of CRISPR‐Cas12a (cpf1) for the Development of a Universal Electrochemical Biosensor

TLDR

An accurate, rapid, and cost‑effective biosensor for disease biomarkers is vital for early‑diagnostic point‑of‑care systems, and the recent discovery of CRISPR type V trans‑cleavage makes CRISPR a potential high‑accuracy bio‑recognition tool. The study reports a CRISPR‑Cas12a (Cpf1) based electrochemical biosensor (E‑CRISPR) that is more cost‑effective and portable than optical trans‑detection, and designs an aptamer‑based E‑CRISPR cascade for detecting TGF‑β1 protein in clinical.

Abstract

An accurate, rapid, and cost-effective biosensor for the quantification of disease biomarkers is vital for the development of early-diagnostic point-of-care systems. The recent discovery of the trans-cleavage property of CRISPR type V effectors makes CRISPR a potential high-accuracy bio-recognition tool. Herein, a CRISPR-Cas12a (cpf1) based electrochemical biosensor (E-CRISPR) is reported, which is more cost-effective and portable than optical-transduction-based biosensors. Through optimizing the in vitro trans-cleavage activity of Cas12a, E-CRIPSR was used to detect viral nucleic acids, including human papillomavirus 16 (HPV-16) and parvovirus B19 (PB-19), with a picomolar sensitivity. An aptamer-based E-CRISPR cascade was further designed for the detection of transforming growth factor β1 (TGF-β1) protein in clinical samples. As demonstrated, E-CRISPR could enable the development of portable, accurate, and cost-effective point-of-care diagnostic systems.