A single-cell RNA-sequencing time course of liver colonisation by colorectal cancer metastases

Murine colorectal cancer cells were injected into the portal vein of NOD.Cg-PrkdcSCID Il2rgtm1Wjl/SzJ mice. Liver metastatic cells were harvested at specific time points for single-cell RNA-sequencing (SORT-seq). Overall design: Lgr5- or Lgr5+ cells were collected from RFP+ murine colorectal cancer organoids (genetic background VillinCre-ERT2; ApcFL/FL; KrasLSL-G12D/+; Tp53KO/KO; R26R-Confetti; Lgr5DTR-eGFP) by FACS and injected into the portal vein of recipient mice. After 1, 7, 13, and 27 days, livers were perfused with a collagenase cocktail and RFP+ metastatic cells were isolated by FACS and analysed by single-cell RNA-seq.