Functional genetic dropout screens and in vivo validation of candidate therapeutic targets using mouse mammary tumoroids

We have previously developed 3D tumoroids derived from genetically engineered mouse models (GEMMs) of BRCA1/2-deficient breast cancer. Here, we describe how to genetically modify tumoroids, use them for functional genetic dropout screens, and carry out orthotopic transplantation of modified tumoroids. The purpose of this protocol is to screen for therapeutic targets and allow rapid and straightforward in vivo validation of the candidate targets. For complete details on the use and execution of this protocol, please refer to Paes Dias et al. (2021b). Keywords: Cancer; Cell Biology; High Throughput Screening; Model Organisms; Molecular Biology; Organoids; Sequencing.