Act-Seq

Act-Seq

Act-Seq minimizes artificially induced transcriptional perturbations common in scRNA-Seq workflows. This is achieved by immediately treating dissociated cells with actinomycin D (actD), a transcription inhibitor of all three eukaryotic RNA polymerases. Authors showed that actD treatment is especially important in inhibiting dissociation-induced immediate-early genes (IEGs) activation that are often seen in scRNA-Seq experiments using conventional dissociation methods. scRNA-Seq of individual cells are assayed using the Drop-Seq method. This method is ideal for interrogating acute transcriptional changes upon external stimuli.

Published: October 2017

Pros:

• Ideal for cell types with low RNA content or where cytoplasmic RNA capture is crucial
• Eliminates activation of gene expression induced by cell dissociation
• Not reliant on flow-sorting of nuclei after immunostaining

Cons:

• Requires custom microfluidics system (Drop-Seq)

• Act-Seq: Wu YE, Pan L, Zuo Y, Li X and Hong W. Detecting Activated Cell Populations Using Single-Cell RNA-Seq. Neuron. 2017;96:313-329.e316

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