Abstract
Organoids are 3D ex vivo cell aggregates derived from primary tissue and shown to closely recapitulate tissue homeostasis. Organoids deliver certain advantages compared to 2D cell lines and mouse models, especially in drug-screening studies and translational research projects. The application of organoids in the research field is fast-emerging and new techniques for organoid manipulation are constantly developing. Despite recent advances, RNA-seq-based drug-screening platforms in organoids are not yet established. Here, we provide a detailed protocol for performing TORNADO-seq, a targeted RNA-seq-based drug-screening method in organoids. Analyzing complex phenotypes with a large number of carefully selected read-outs allows to directly classify and group drugs even without structural similarity or overlapping mode of actions from prior knowledge. Our assay principle combines cost-effectiveness and sensitive detection of multiple cell identities, signaling pathways, and key drivers of cellular phenotypes and can be applied to many systems where this new form of high-content screening can provide information not obtainable otherwise.
Key words
- Targeted RNA-seq organoids
- High-throughput screening
- High-content screening
- NGS-based screening in organoids
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Norkin, M., Huelsken, J. (2023). TORNADO-seq: A Protocol for High-Throughput Targeted RNA-seq-Based Drug Screening in Organoids. In: Ordóñez-Morán, P. (eds) Intestinal Differentiated Cells. Methods in Molecular Biology, vol 2650. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3076-1_6
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DOI: https://doi.org/10.1007/978-1-0716-3076-1_6
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