Temporal dynamics of collateral RNA cleavage by LbuCas13a in human cells
We have found that LbuCas13a exhibits collateral RNA cleavage activity in human cells after activation by the target RNA. Here we use total RNA sequencing to explore which RNAs are depleted by the collateral activity of LbuCas13a. ERCC spike-in RNAs were used for normalization between samples. This revealed near global depletion of mRNAs at 50 and 200 minutes after Cas13 protein and guide delivery. Mitochondrial and nuclear transcripts were not affected. Overall design: Total RNA sequencing of HAP1-dEGFP cells with ERCC spike-in control RNAs. Cells were either left untreated, transfected with an empty iTOP transfection, transfected with LbuCas13a and a non-targeting gRNA by iTOP, or transfected with LbuCas13a and a dEGFP targeting gRNA by iTOP. Cells were harvested at 50, 200 and 1440 minutes after transfection.