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Will seed sibling controls always correctly identify phenotypes due to off-target effects?

No single approach should be relied upon to answer the important question of whether a hit in a siRNA screen is real or not. However, if there are multiple seed siblings that give a phenotype and do not silence the target gene of the experimental siRNA, it is highly likely to be due to an off-target. Similarly, the lack of phenotype with seed siblings will not guarantee that a hit is real, but it certainly will raise the confidence level of that siRNA/gene target as a real hit.