Work

Protocol for first-hit oncogenic organoid modeling and lentiviral functional screening

Year: 2026

Type: article

Abstract: We present a protocol for modeling early tumorigenesis using air-liquid interface (ALI) organoids derived from oral, esophageal, and lung epithelia bearing first-hit oncogenic mutations. We describe steps for identifying candidate amplified oncogenes through outlier analysis, establishing ALI organoids, and inducing oncogenic activation. We then perform pooled lentiviral open reading frame (ORF) library delivery, selection, and barcode sequencing for functional screening. This approach enables validation of candidate drivers of neoplastic transformations, addressing a gap between in vitro and in vivo tumorigenesis studies. For complete details on the use and execution of this protocol, please refer to Salahudeen et al. 1 1. Salahudeen, A.A. ∙ Seoane, J.A. ∙ Yuki, K. ... Functional screening of amplification outlier oncogenes in organoid models of early tumorigenesis Cell Rep. 2023; 42 , 113355 Google Scholar

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104625

Authors: Farah Yousef, Jaramys Mosley, Olga Karginova, Jose A. Seoane, Ameen A. Salahudeen

Institutions: University of Illinois Chicago, Vall d'Hebron Institute of Oncology

Language: English

Topic: Cancer Genomics and Diagnostics

Subfield: Cancer Research

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550