PT  - JOURNAL ARTICLE
AU  - William Putzbach
AU  - Ashley Haluck-Kangas
AU  - Quan Q. Gao
AU  - Aishe A. Sarshad
AU  - Elizabeth T. Bartom
AU  - Austin Stults
AU  - Abdul S. Qadir
AU  - Markus Hafner
AU  - Marcus E. Peter
TI  - CD95L mRNA is toxic to cells
AID  - 10.1101/330324
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 330324
4099  - http://biorxiv.org/content/early/2018/05/24/330324.short
4100  - http://biorxiv.org/content/early/2018/05/24/330324.full
AB  - CD95/Fas ligand binds to the death receptor CD95/Fas to induce apoptosis in sensitive cells. We previously reported the CD95L mRNA is enriched in sequences that, when converted to si/shRNAs, are toxic to cells (Putzbach et al., 2017). These si/shRNAs kill all cancer cells through a RNAi off-target effect by targeting critical survival genes. 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 RNAs derived from CD95L are loaded into the RNA induced silencing complex (RISC) and that the RISC is required for the toxicity. Drosha and Dicer knock-out cells are highly sensitive to this toxicity, suggesting that processing of CD95L mRNA into small toxic RNAs is independent of both Dicer and Drosha. The data provide evidence that a higher vertebrate transgene can be processed to RNAi-active small RNAs that elicit cellular responses.