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

Mariana Paes Dias; Sven Rottenberg; Jos Jonkers

doi:10.1016/j.xpro.2022.101132

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

STAR Protoc. 2022 Jan 26;3(1):101132. doi: 10.1016/j.xpro.2022.101132. eCollection 2022 Mar 18.

Authors

Mariana Paes Dias^{1

2}, Sven Rottenberg^{1

3}, Jos Jonkers^{1

2}

Affiliations

¹ Division of Molecular Pathology, the Netherlands Cancer Institute, 1066CX Amsterdam, the Netherlands.
² Oncode Institute, 1066CX Amsterdam, the Netherlands.
³ Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Bern 3012, Switzerland.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Breast Neoplasms* / genetics
Female
Humans
Mice