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Updated: Jun 14, 2026
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A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids
Published on: April 20, 2015
Protocol for first-hit oncogenic organoid modeling and lentiviral functional screening
Farah Yousef1, Jaramys Mosley2, Olga Karginova1
1Department of Medicine, Division of Hematology and Oncology, University of Illinois College of Medicine, Chicago, IL 60612, USA; Department of Biochemistry and Molecular Genetics, University of Illinois College of Medicine, Chicago, IL 60612, USA.
STAR Protocols
|June 12, 2026
Summary
This study introduces a novel protocol for modeling early tumorigenesis using air-liquid interface (ALI) organoids. The method validates oncogenes driving neoplastic transformations, bridging in vitro and in vivo research gaps.
Area of Science:
- Oncology
- Cell Biology
- Genetics
Background:
- Early tumorigenesis modeling is crucial for understanding cancer development.
- Existing methods have limitations in bridging in vitro and in vivo studies.
- Identifying key oncogenes driving neoplastic transformation requires robust experimental systems.
Purpose of the Study:
- To present a protocol for modeling early tumorigenesis using air-liquid interface (ALI) organoids.
- To enable functional screening of candidate oncogenes involved in neoplastic transformation.
- To validate oncogene drivers and address limitations in current tumorigenesis research.
Main Methods:
- Utilizing oral, esophageal, and lung epithelial ALI organoids with initial oncogenic mutations.
- Employing outlier analysis to identify candidate amplified oncogenes.
- Performing pooled lentiviral open reading frame (ORF) library delivery, selection, and barcode sequencing for functional screening.
Main Results:
- The protocol successfully models early tumorigenesis in epithelial organoids.
- Candidate oncogenes driving neoplastic transformations can be identified and validated.
- This approach provides a bridge between in vitro and in vivo tumorigenesis studies.
Conclusions:
- The developed ALI organoid protocol offers a powerful platform for studying early tumorigenesis.
- This method facilitates the validation of oncogenes as drivers of neoplastic transformation.
- The protocol addresses a critical gap in cancer research, enhancing the study of tumorigenesis.
