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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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Updated: Aug 10, 2025

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

A universal and sensitive gene mutation detection method based on CRISPR-Cas12a.

Huajing Wang¹, Ruijie Liu², Kejun Dong³

¹Department of Breast Surgery, Second Hospital of Jilin University, No.218 Ziqiang Street, Nanguan District, Changchun, 130041, China; Institute of Reproductive Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.

Analytica Chimica Acta

|February 10, 2023

Summary

This summary is machine-generated.

This study introduces a novel CRISPR/Cas12a method for highly sensitive and specific detection of single nucleotide mutations, crucial for early cancer diagnosis and treatment. The method achieves 100% accuracy in clinical samples.

Keywords:

CRISPR-Cas12a Gene mutation Sensitivity Specific PCR Universality

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

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

Published on: December 23, 2022

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Single nucleotide mutations are key drivers in cancer development.
Accurate detection of these mutations is vital for clinical oncology.
Existing CRISPR/Cas12a detection methods have limitations in specificity and universality.

Purpose of the Study:

To develop a simple, rapid, and highly sensitive method for single nucleotide mutation detection using CRISPR/Cas12a.
To overcome the PAM sequence restriction and enhance the specificity and sensitivity of CRISPR/Cas12a.
To validate the clinical applicability of the developed method for cancer mutation detection.

Main Methods:

Development of a novel CRISPR/Cas12a-based detection system.
Integration with specific PCR to overcome PAM sequence limitations.
Detection of clinically significant mutations (PTEN R130Q, BRAF V600E, TP53 R248W).

Main Results:

Achieved a detection limit of 0.1% for single nucleotide mutations.
Demonstrated significantly improved specificity and sensitivity compared to existing methods.
Validated with 100% accuracy on 10 clinical samples for TP53 R248W mutation.

Conclusions:

The developed specific PCR combined with CRISPR/Cas12a method is simple, rapid, universal, and highly sensitive.
This method shows significant promise for clinical cancer diagnosis.
Overcoming PAM sequence restrictions enhances the clinical utility of CRISPR/Cas12a for mutation detection.