Efficient and Stable Subcellular Protein Labeling in Leishmania mexicana Using a Re-Engineered mNeonGreen Integration
Tianyu Lei1, Mengtao Yu1, Panjing Lv1
1Department of Pathogen Biology, School of Basic Medicine, Tongji Medical College, State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Huazhong University of Science and Technology, Wuhan 430030, China.
Pathogens (Basel, Switzerland)
|May 4, 2026
Summary
Researchers developed a new molecular toolbox for Leishmania mexicana, using a bright mNeonGreen fluorophore for stable protein localization. This tool enhances high-resolution imaging of parasite organelle dynamics.
Area of Science:
- Parasitology
- Molecular Biology
- Cell Biology
Background:
- Leishmania mexicana is a key model for trypanosomatid research.
- Existing fluorescent tools lack stability and intensity for precise subcellular localization.
Purpose of the Study:
- To develop a stable, high-intensity fluorescent reporter system for Leishmania mexicana.
- To enable precise subcellular protein localization and high-resolution imaging.
Main Methods:
- Re-engineered the pLEXSY-hyg2.1 vector to express mNeonGreen (mNG).
- Utilized modular cloning for N-terminal protein tagging.
- Integrated the construct into the 18S rRNA locus for stable expression.
Main Results:
- Created a versatile molecular toolbox with a bright, photostable mNG fluorophore.
- Demonstrated stable, constitutive expression of tagged proteins.
- Generated a proof-of-concept transgenic line with specific flagellar localization.
Conclusions:
- The integration-based system provides an efficient and stable platform for Leishmania protein visualization.
- This tool simplifies reporter strain generation for high-resolution imaging and functional genomics.
