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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/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

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A chemical-enhanced system for CRISPR-Based nucleic acid detection.

Zihan Li¹, Wenchang Zhao¹, Shixin Ma¹

¹College of Life and Health Sciences, Northeastern University, Shenyang, 110819, People's Republic of China; Key Laboratory of Data Analytics and Optimization for Smart Industry (Northeastern University), Ministry of Education, Shenyang, 110819, People's Republic of China.

Biosensors & Bioelectronics

|July 16, 2021

Summary

Chemical Enhanced CRISPR Detection (CECRID) improves viral pathogen detection by optimizing reaction chemistry. L-proline enhances CRISPR-based nucleic acid detection sensitivity for point-of-care diagnostics.

Keywords:

COVID-19 CRISPR Chemical Detection Diagnostics SARS-CoV-2

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Published on: April 25, 2025

Area of Science:

Molecular Biology
Biotechnology
Diagnostic Assays

Background:

CRISPR-based nucleic acid detection systems offer potential for point-of-care viral pathogen testing, particularly relevant during the COVID-19 pandemic.
Optimization of reaction chemistry is crucial for enhancing the performance of these diagnostic systems.

Purpose of the Study:

To optimize key reaction chemistry parameters for CRISPR-based nucleic acid detection.
To develop an enhanced CRISPR detection system, termed Chemical Enhanced CRISPR Detection (CECRID), for improved sensitivity and robustness.

Main Methods:

Optimized buffer conditions and substrate range for Cas12a/Cas13a-based signal detection.
Investigated the role of bovine serum albumin and chemical additives, specifically L-proline, in enhancing CRISPR detection.
Assessed the impact of L-proline on isothermal amplification methods (LAMP and RPA).
Validated the CECRID system using SARS-CoV-2 pseudovirus with fluorescence and lateral flow strip readouts.

Main Results:

Bovine serum albumin was found to be crucial for enhancing the trans-cleavage activity of Cas12a/Cas13a effectors.
L-proline significantly enhanced Cas12a/Cas13a detection capability and specific target amplification in LAMP and RPA.
CECRID demonstrated enhanced detection sensitivity compared to methods without chemical additives when tested with SARS-CoV-2 pseudovirus.

Conclusions:

The developed Chemical Enhanced CRISPR Detection (CECRID) system significantly improves detection power and robustness.
CECRID provides a foundation for developing enhanced reagent formulations and test kits for practical CRISPR-based diagnostics.
This optimization strategy holds promise for advancing point-of-care diagnostic applications for viral pathogens.