Similar seed effects in independent siRNA screens
A 2013 study on Parkin translocation used genome-wide siRNA libraries from Ambion (single Silencer Select siRNAs) and Dharmacon (pools of 4 siGENOME siRNAs).
Low hit validation rate for Dharmacon siGENOME screens
Good experimental design is important when validating hits from RNAi screens.
Pooling only 4 siRNAs increases off-target effects
Low-complexity pools (with 4 siRNAs per gene) should thus lead to overall stronger off-target effects than single siRNAs.
Performing target validation well
This blogpost describes issues encountered in target validation and how to safeguard against poor reproducibility in RNAi experiments.
Low complexity pooling does not prevent siRNA off-targets
Low-complexity siRNA pooling (e.g. Dharmacon siGENOME SMARTpools) does not prevent siRNA off-targets. It may in fact exacerbate off-target effects. Only high-complexity pooling (siPOOLs) can reliably ensure on-target phenotypes.
What is the probability of an siRNA off-target phenotype?
Conventional siRNAs have a high probability of giving off-target phenotypes. siRNA off-target effects can be reduced by using more specific reagents or narrowing the assay focus (to reduce the number of relevant genes).
Correcting seed-based off-target effects in RNAi screens
Unlocking RNAi screening potential by correcting seed-based off-target effects. While seed correction methods yield minor improvements, challenges persist in interpreting strongest hits. The quest for precise reagents, like high-complexity siPOOLs, remains crucial for reliable results.
Novel anti-cancer mechanism identified by shRNA/siRNA off-target effects
Unveiling a novel anti-cancer mechanism via siRNA/shRNA off-target effects. Putzbach et al.'s study exposes survival genes in cancer cells, shedding light on CD95/CD95L-induced cell death and the role of miRNA-like activity in off-target effects
Making sense of siGENOME deconvolution
Deconvolution of Dharmacon siGENOME pools shows surprising results, with low correlation to target genes and significant seed effects.
The final RNAiL?
Explore the debate over RNAi's viability post-CRISPR revelations. Lin et al.'s findings challenge RNAi's efficacy, but nuances suggest RNAi still has value, albeit with precautions against off-target effects
