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Updated: Sep 16, 2025

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

Published on: April 25, 2022

Regulating cleavage activity and enabling microRNA detection with split sgRNA in Cas12b.

Jiaqi Wang¹, Xiaofang Ye¹, Yuanfang Liu¹

¹Guangdong Provincial Key Laboratory of Digestive Cancer Research, Digestive Diseases Center, Scientific Research Center, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, Guangdong, China.

Nature Communications

|July 10, 2025

Summary

This summary is machine-generated.

A novel split single guide RNA (sgRNA) strategy enhances CRISPR-Cas12b diagnostics. This adaptable system allows precise nucleic acid detection, including microRNAs, with improved regulation and simplified protocols for clinical applications.

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Area of Science:

Molecular Biology
Biotechnology
Nucleic Acid Detection

Background:

CRISPR-Cas12b systems offer advanced molecular diagnostics.
Current limitations include reliance on long single guide RNAs (sgRNAs) >100 nt, hindering precise control.
Need for adaptable and regulatable CRISPR systems for diverse applications.

Purpose of the Study:

To develop a split sgRNA strategy for CRISPR-Cas12b to enable precise regulation and broader applications.
To create a versatile Cas12b system using universal components and a customizable Spacer.
To introduce dynamic control mechanisms for Cas12b activity.

Main Methods:

Designed a split sgRNA system for Cas12b with universal components and a replaceable Spacer.
Utilized glyoxal labeling and photo-cleavable linkers for dynamic Cas12b activity modulation.
Validated detection of Epstein-Barr virus in clinical samples and microRNAs without amplification.

Main Results:

The split sgRNA strategy allows simple target detection by replacing the Spacer.
Glyoxal labeling and photo-cleavable linkers provide temperature- and UV-mediated dynamic regulation.
Achieved sensitive detection of Epstein-Barr virus in plasma, comparable to qPCR.
Demonstrated direct microRNA detection without reverse transcription or amplification.
Consistent results with RT-qPCR in clinical samples (healthy individuals, colorectal cancer patients).

Conclusions:

The split sgRNA strategy significantly enhances CRISPR-Cas12b systems for molecular diagnostics.
This approach offers a simplified, highly adaptable, and precisely regulated method for clinical nucleic acid analysis.
The system shows promise for direct detection of biomarkers like microRNAs.