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CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

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Synthetic Protein-to-DNA Input Exchange for Protease Activity Detection Using CRISPR-Cas12a.

Luca Capelli¹, Federica Pedrini¹, Andrea C Di Pede²

¹Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parco Area Delle Scienze 17/A, 43124 Parma, Italy.

Analytical Chemistry

|November 14, 2024

Summary

A new method detects matrix metalloproteinase 2 (MMP2), a cancer biomarker, using a chemical translator and CRISPR-Cas12a signal amplification. This activity-based assay achieves sensitive MMP2 detection in the low picomolar range.

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Substrate Generation for Endonucleases of CRISPR/Cas Systems

Substrate Generation for Endonucleases of CRISPR/Cas Systems

Published on: September 8, 2012

Area of Science:

Biochemistry
Molecular Biology
Biosensing

Background:

Matrix metalloproteinase 2 (MMP2) is a key protease biomarker in cancer progression.
Sensitive and specific detection of MMP2 is crucial for early cancer diagnosis and monitoring.
Existing detection methods may lack the sensitivity or specificity required for clinical applications.

Purpose of the Study:

To develop a novel activity-based detection strategy for MMP2.
To integrate MMP2 activity with CRISPR-Cas12a for enhanced signal amplification.
To establish a sensitive and specific assay for MMP2 detection.

Main Methods:

Designed a chemical translator (peptide-PNA conjugate) acting as an MMP2 substrate and nucleic acid output.
Immobilized the translator on magnetic beads for an activity-based assay.
Utilized CRISPR-Cas12a system with a guide RNA and FRET-labeled DNA reporters for signal amplification.

Main Results:

The assay demonstrated high sensitivity for MMP2 detection, with a limit of detection of 72 pg/mL (low picomolar range).
The system effectively leveraged MMP2 proteolytic activity for signal generation.
CRISPR-Cas12a amplification significantly enhanced the fluorescence signal.

Conclusions:

The developed strategy provides a sensitive and specific method for MMP2 detection.
This approach offers new design principles for CRISPR-Cas-based biosensors.
The findings have implications for developing advanced diagnostic tools for cancer biomarkers.