|Publication Type||Journal Article|
|Year of Publication||2015|
|Authors||Singh, S, Wu, X, Ljosa, V, Bray, M-A, Piccioni, F, Root, DE, Doench, JG, Boehm, JS, Carpenter, AE|
|Keywords||Cell Line, Gene Expression Profiling, Gene Knockdown Techniques, Humans, RNA Interference, RNA, Small Interfering|
RNA interference and morphological profiling-the measurement of thousands of phenotypes from individual cells by microscopy and image analysis-are a potentially powerful combination. We show that morphological profiles of RNAi-induced knockdown using the Cell Painting assay are in fact highly sensitive and reproducible. However, we find that the magnitude and prevalence of off-target effects via the RNAi seed-based mechanism make morphological profiles of RNAi reagents targeting the same gene look no more similar than reagents targeting different genes. Pairs of RNAi reagents that share the same seed sequence produce image-based profiles that are much more similar to each other than profiles from pairs designed to target the same gene, a phenomenon previously observed in small-scale gene-expression profiling experiments. Various strategies have been used to enrich on-target versus off-target effects in the context of RNAi screening where a narrow set of phenotypes are measured, mostly based on comparing multiple sequences targeting the same gene; however, new approaches will be needed to make RNAi morphological profiling (that is, comparing multi-dimensional phenotypes) viable. We have shared our raw data and computational pipelines to facilitate research.
|Alternate Journal||PLoS ONE|
|PubMed Central ID||PMC4511418|