Protocol for the characterization of the pancreatic tumor microenvironment using organoid-derived mouse models and single-nuclei RNA sequencing

STAR Protoc. 2024 Sep 20;5(3):103203. doi: 10.1016/j.xpro.2024.103203. Epub 2024 Jul 24.

Abstract

Single-nuclei RNA sequencing (snRNA-seq) allows for obtaining gene expression profiles from frozen or hard-to-dissociate tissues at the single-nuclei level. Here, we describe a protocol to obtain snRNA-seq data of pancreatic tumors from orthotopically grafted organoid-derived mouse models. We provide details on the establishment of these mouse models, the isolation of single nuclei from pancreatic tumors, and the analysis of the snRNA-seq datasets. For complete details on the use and execution of this protocol, please refer to Mucciolo et al.1.

Keywords: Cancer; Genomics; Model Organisms; Organoids; RNAseq; Sequence analysis; Sequencing.

MeSH terms

  • Animals
  • Humans
  • Mice
  • Organoids* / metabolism
  • Organoids* / pathology
  • Pancreatic Neoplasms* / genetics
  • Pancreatic Neoplasms* / pathology
  • RNA, Small Nuclear / genetics
  • RNA, Small Nuclear / metabolism
  • Sequence Analysis, RNA* / methods
  • Tumor Microenvironment* / genetics

Substances

  • RNA, Small Nuclear