Related Experiment Video
Updated: May 20, 2025
In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
Published on: March 29, 2019
Nuclear-Targeted Material Enabled Intranuclear MicroRNA Imaging for Tracking Gene Editing Process
Jiayan Wu1,2, Meng Meng2, Zhaopei Guo2
1State Key Laboratory of Physical Chemistry of Solid Surfaces, College of Chemistry and Chemical Engineering, Innovation Laboratory for Sciences and Technologies of Energy Materials of Fujian Province (IKKEM), Xiamen University, Xiamen, 361005, China.
Abstract:
Gene editing technology based on clustered regularly interspaced short palindromic repeats/associated protein (CRISPR/Cas) systems serves as an efficient tool in cancer therapy. Tracking the gene editing process can help identify the progress of cancer treatment. However, existing techniques for monitoring the gene editing process rely on lysed cells, which can not reflect the dynamic changes of nucleic acid in living cells. It urgently needs in situ and real-time imaging technologies to track the gene editing process at a living single-cell level more effectively and precisely. Here, we reported a highly efficient nuclear-targeted material, phenylboronic acid modified linear PEI (LPBA), for loading gene editing plasmids and fluorescent probes to track gene editing processes of microRNA. Based on LPBA, we achieved efficient intranuclear microRNA imaging at the living cell level, reaching 32.4-fold higher than the linear PEI (LPEI) delivery system, which facilitated further sensitive monitoring of the gene editing process both in living cells and in vivo. Meanwhile, this efficient gene-editing and real-time detection technique could be extended to screening effective gene-editing plasmids. Such LPBA-based imaging technology extended the imaging area of microRNA and offered new insight in the field of gene editing and nucleic acid detection.
Insights
Researchers developed a new material (LPBA) for tracking gene editing in living cells. This breakthrough enables real-time monitoring of microRNA gene editing, improving cancer therapy insights and plasmid screening.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Clustered regularly interspaced short palindromic repeats/associated protein (CRISPR/Cas) systems are vital for cancer therapy.
- Current methods for tracking gene editing use lysed cells, failing to capture dynamic changes in living cells.
- There is a critical need for in situ, real-time imaging to monitor gene editing at the single-cell level.
Purpose of the Study:
- To develop a novel nuclear-targeted material for efficient gene editing plasmid delivery and tracking.
- To achieve in situ and real-time imaging of microRNA gene editing processes in living cells.
- To enable sensitive monitoring of gene editing in vitro and in vivo and facilitate screening of effective gene-editing plasmids.
Main Methods:
- A phenylboronic acid modified linear polyethyleneimine (LPBA) material was synthesized for nuclear targeting.
- LPBA was used to load gene editing plasmids and fluorescent probes for microRNA imaging.
- The LPBA system's efficiency was compared to the linear polyethyleneimine (LPEI) delivery system in living cells and in vivo.
Main Results:
- LPBA demonstrated highly efficient nuclear targeting and loading of gene editing plasmids and probes.
- Intranuclear microRNA imaging in living cells was achieved with LPBA, showing a 32.4-fold increase compared to LPEI.
- The LPBA system facilitated sensitive, real-time monitoring of gene editing processes in both living cells and in vivo.
Conclusions:
- The developed LPBA material enables efficient intranuclear imaging of microRNA gene editing in living cells.
- This technique provides a powerful tool for real-time monitoring of gene editing and screening of gene-editing plasmids.
- The LPBA-based imaging technology offers new insights into gene editing and nucleic acid detection for cancer therapy.
Related Concept Videos
02:15Experimental RNAi
01:22MicroRNAs