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Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications
Jasmin Khateeb1,2, Jady Liang1,3, Yuchong Li1,4
1Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Unity Health Toronto, Toronto, ON, Canada.
Communications Biology
|February 3, 2025
Summary
Researchers improved gene delivery into human lung organoids using induced pluripotent stem cells (iPSCs). A new method enhances lentiviral vector transduction efficiency for better lung research and disease modeling.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Molecular Biology
Background:
- Human induced pluripotent stem cell (iPSC)-derived lung organoids are valuable for studying lung development and disease.
- Lentiviral vectors (LVVs) are effective for stable gene expression but challenging to deliver into complex organoid structures.
Purpose of the Study:
- To optimize lentiviral vector (LVV) transduction efficiency in human iPSC-derived lung organoids.
- To develop a reliable method for genetic engineering of lung organoids for research applications.
Main Methods:
- Physically disrupting organoids to increase surface area.
- Employing spinoculation to enhance viral particle binding and entry.
- Utilizing an optimized culture medium to support transduction.
- Validating transduction efficiency using single-cell RNA sequencing (scRNA-seq) and molecular assays.
Main Results:
- The optimized protocol significantly enhanced LVV transduction efficiency in lung organoids.
- Successful gene and protein level validation confirmed the method's efficacy.
- The technique improved the ability to genetically modify iPSC-derived lung organoids.
Conclusions:
- A novel, optimized transduction method enhances LVV delivery into human iPSC-derived lung organoids.
- This protocol facilitates genetic manipulation for advanced lung research.
- The findings provide a valuable tool for studying lung development and disease mechanisms in vitro.
