Abstract
Over 80% of multiple-tested siRNAs and shRNAs targeting CD95 or CD95 ligand (CD95L) induce a form of cell death characterized by simultaneous activation of multiple cell death pathways preferentially killing transformed and cancer stem cells. We now show these si/shRNAs kill cancer cells through canonical RNAi by targeting the 3’UTR of critical survival genes in a unique form of off-target effect we call DISE (death induced by survival gene elimination). Drosha and Dicer-deficient cells, devoid of most miRNAs, are hypersensitive to DISE, suggesting cellular miRNAs protect cells from this form of cell death. By testing 4666 shRNAs derived from the CD95 and CD95L mRNA sequences and an unrelated control gene, Venus, we have identified many toxic sequences-most of them located in the open reading frame of CD95L. We propose that specific toxic RNAi-active sequences present in the genome can kill cancer cells.
| Original language | English (US) |
|---|---|
| Article number | e29702 |
| Journal | eLife |
| Volume | 6 |
| DOIs | |
| State | Published - Oct 24 2017 |
Funding
We are grateful to Lindsay Stolzenburg and Ann Harris for helping to set up the CRISPR/Cas9 gene editing method and to Matthew Schipma for computational support. We would like to thank the Gene Editing and Screening Core, at Memorial Sloan Kettering in New York City, NY, for RNAi reagents and services. MH was supported by the Intramural Research Program of NIAMS. AAS. acknowledges support by the Swedish Research Council postdoctoral fellowship. This work was funded by training grants T32CA070085 (to MP) and T32CA009560 (to WP) R50CA211271 (to JCZ), and R35CA197450 (to MEP).
ASJC Scopus subject areas
- General Neuroscience
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology
