Project description:CD95/Fas ligand binds to the death receptor CD95 to induce apoptosis in sensitive cells. We previously reported that CD95L mRNA is enriched in sequences that, when converted to si/shRNAs, kill all cancer cells by targeting critical survival genes (Putzbach et al., 2017). We now report expression of full-length CD95L mRNA itself is highly toxic to cells and induces a similar form of cell death. We demonstrate that small (s)RNAs derived from CD95L are loaded into the RNA induced silencing complex (RISC) which is required for the toxicity and processing of CD95L mRNA into sRNAs is independent of both Dicer and Drosha. We provide evidence that in addition to the CD95L transgene a number of endogenous protein coding genes involved in regulating protein translation, particularly under low miRNA conditions, can be processed to sRNAs and loaded into the RISC suggesting a new level of cell fate regulation involving RNAi.
Project description:Analysis of RISC bound short (s)RNAs in a HCT116 Drosha CD95 d.k.o. cell line expressing pLenti-CD95L and various CD95L mutant constructs.
Project description:Analysis of RISC bound short (s)RNAs in a HCT116 wild-type, Drosha k.o., and Ago 1/2/3 k.o. cells expressing pLenti-CD95L NP or pLenti empty vector.
Project description:Analysis of RISC bound short (s)RNAs in a HCT116 Drosha CD95 d.k.o. cell line expressing pLenti-CD95L and various CD95L mutant constructs.
Project description:Analysis of RISC bound short (s)RNAs in a HCT116 wild-type, Drosha k.o., and Dicer k.o. cells expressing pLenti-CD95L NP or pLenti empty vector.
