Identification of DISE-inducing shRNAs by monitoring cellular responses

Cell Cycle
Monal Patel, Marcus E Peter

Abstract

Off-target effects (OTE) are an undesired side effect of RNA interference (RNAi) caused by partial complementarity between the targeting siRNA and mRNAs other than the gene to be silenced. The death receptor CD95 and its ligand CD95L contain multiple sequences that when expressed as either si- or shRNAs kill cancer cells through a defined OTE that targets critical survival genes. Death induced by survival gene elimination (DISE) is characterized by specific morphological changes such as elongated cell shapes, senescence-like enlarged cells, appearance of large intracellular vesicles, release of mitochondrial ROS followed by activation of caspase-2, and induction of a necrotic form of mitotic catastrophe. Using genome-wide shRNA lethality screens with eight different cancer cell lines, we recently identified 651 genes as critical for the survival of cancer cells. To determine whether the toxic shRNAs targeting these 651 genes contained shRNAs that kill cancer cell through DISE rather than by silencing their respective target genes, we tested all shRNAs in the TRC library derived from a subset of these genes targeting tumor suppressors (TS). We now report that only by monitoring the responses of cancer cells following expression ...Continue Reading

References

Oct 26, 2000·Nature·P H Krammer
Feb 7, 2001·Genes & Development·S M ElbashirT Tuschl
Apr 1, 2005·Nature Structural & Molecular Biology·Fabiola V RivasLeemor Joshua-Tor
Aug 11, 2005·Nucleic Acids Research·Xiaoyu LinYu Shen
Feb 21, 2006·Nature Methods·Amanda BirminghamAnastasia Khvorova
Feb 26, 2009·Cell·Richard W Carthew, Erik J Sontheimer
Aug 18, 2009·Nature Structural & Molecular Biology·Tomoko KawamataYukihide Tomari
Dec 8, 2009·Nature Structural & Molecular Biology·Mayuko YodaYukihide Tomari
Aug 28, 2012·Chemistry & Biology·Glen F Deleavey, Masad J Damha
Oct 16, 2012·Nucleic Acids Research·Min ZhaoZhongming Zhao
Feb 26, 2013·Methods in Molecular Biology·Sebastian Petri, Gunter Meister
Jul 3, 2013·Cold Spring Harbor Protocols·Kenneth ChangGregory J Hannon
Dec 18, 2013·Cell Reports·Christof FellmannJohannes Zuber
Mar 25, 2014·Cell Reports·Abbas HadjiMarcus E Peter
Nov 5, 2014·Nature Communications·Paolo CeppiMarcus E Peter
Nov 6, 2014·Genome Research·Patrick EssletzbichlerTilmann Bürckstümmer
May 10, 2016·Nature Biotechnology·David W MorgensMichael C Bassik
Nov 22, 2017·Oncotarget·Andrea E MurmannMarcus E Peter

❮ Previous
Next ❯

Citations

Oct 31, 2018·Nature Communications·Quan Q GaoMarcus E Peter
Oct 17, 2018·ELife·Will PutzbachMarcus E Peter
Apr 16, 2021·Journal of Pharmaceutical and Biomedical Analysis·Sinem Nalbantoglu, Abdullah Karadag
Apr 15, 2021·Molecular Therapy. Nucleic Acids·Joshua M CorbinMaria J Ruiz-Echevarría
Dec 12, 2021·Journal of Experimental & Clinical Cancer Research : CR·Ashley Haluck-KangasMarcus E Peter

❮ Previous
Next ❯

Methods Mentioned

BETA
transfecting
RNA Seq
flow cytometry
Protein Assay

Software Mentioned

IncuCyte
STATA1C

Related Concepts

Related Feeds

Apoptotic Caspases

Apoptotic caspases belong to the protease enzyme family and are known to play an essential role in inflammation and programmed cell death. Here is the latest research.