Effect of Trp53 knockout on fallopian tube epithelial organoids

We established murine fallopian tube epithelial organoids from a B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J mouse. Subsequently, we knocked out Trp53 by introducing sgRNA into the organoids, nutlin-3 selection, and single-organoid cloning. The Trp53-knocked organoids grew faster than the normal organoids. We also analyzed the transcriptomic differences caused by Trp53-knockout by RNA-sequencing and gene set enrichment analysis (GSEA), which indicated that Trp53-knockout reduced cilium-related gene expression. In human HGSC precancerous lesions, such as serous tubal intraepithelial carcinoma (STIC), differentiation to ciliated cells has been reported to be down-regulated; therefore, the genetic manipulation was supposed to mimic the process of carcinogenesis of HGSC. To investigate the biological changes caused by TRP53 dysfunction in fallopian tube epithelium, we established murine fallopian tube epithelial organoids and knocked out Trp53. A control sample was normal fallopian tube epithelial organoids established from a B6J.129(B6N)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J mouse.