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. Overall design: 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.