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What level of activation should I expect to see in my target gene when using CRISPRa?

The level of gene activation depends on the basal expression of the gene, and is therefore cell-line specific. For genes that are not expressed or expressed at a low level, we generally see high fold activation by RTqPCR (100 to 10,000 fold over untreated cells), but for genes that are expressed at a high level, CRISPRa causes lowerfold activation (2 -100 fold). Therefore, knowing the level of expression of the gene of interest in your cells can help you gauge the expectation for the fold activation and help in the hit identification and validation.