Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA

Abstract

<p>A generic sensing strategy that integrates CRISPR-Cas12a with a DNA circuit is proposed for amplified detection of microRNA.</p>

A pH-engineering regenerative DNA tetrahedron ECL biosensor for the assay of SARS-CoV-2 RdRp gene based on CRISPR/Cas12a trans-activity

“…With the single-stranded DNA Trans-cleavage activity of the Cas12a enzyme, combined with various isothermal nucleic acid amplification techniques, a series of CRISPR-Cas molecular diagnostic platforms based on fluorescent signal output have been developed [9] . Among the CRISPR-Cas effector family, Cas12 is an RNA-directed DNase that belongs to the class II V-A system, which induces the division of arbitrary single-stranded DNA (ssDNA) upon target recognition, which leads to the degradation of the ssDNA reporter and the release of a fluorescent signal at the division site, which can be detected by a portable method [10] , [11] , [12] . Although optical transduction-based CRISPR-Cas molecular diagnostics have been widely used for nucleic acids [13] , small molecules [14] and metal ions [15] , considering that these strategies need to be done on expensive and complex devices, there is a need to extend the CRISPR/Cas system to the field of solid-load-free electrochemical biosensing.…”

Advances in the Integration of Nucleic Acid Nanotechnology into CRISPR-Cas System

“…[71]. Copyright 2020, The American Association for the Advancement of Science work suggested that the integration of nucleic acid circuit and CRISPR-Cas systems could be an effective strategy to develop sensitive CRISPR-based bioassays [70]. Therefore, the combination of nucleic acid nanotechnology may offer an alternative paradigm for the development and application of powerful CRISPR-based molecular diagnostic methods.…”

“…8b) and biomarkers [68,69]. By employing a DNA circuit for signal conversion, our group described the CRISPR-based sensitive and cost-effective diagnostics method for miRNA detection [70]. This assay can achieve sub-femolar sensitivity for the analysis of different miRNA biomarkers.…”

CRISPR‐Cas12a System for Biosensing and Gene Regulation

“…Therefore, Cas12a has recently been widely expanded as a diagnostic tool for detecting nucleic acids with high sensitivity in vitro [11,25] . To achieve fluorescent detection of low‐abundance nucleic acid targets, it is often necessary to combine some nucleic acid amplification strategies, including polymerase chain reaction (PCR), [23,26] recombinase polymerase amplification (RPA), [9b,c,10a,c,d,f,12,32] loop‐mediated isothermal amplification (LAMP), [10b,e,27,29,33] enhanced strand displacement amplification (E‐SDA), [34] rolling circle amplification (RCA), [35] or catalytic hairpin assembly (CHA) [28] (Table 1).…”