tRNA anticodon cleavage by target-activated CRISPR-Cas13a effector

In this study, we conducted an analysis of RNA cleavage products mediated
by the Type VI CRISPR-Cas system from the bacterium Leptotrichia shahii.
Previous research has demonstrated that the LshCas13a effector protein,
when loaded with CRISPR-RNA, exhibits collateral RNase activity upon
recognizing the target transcript. To identify the products resulting from
RNA cleavage mediated by the activated Type VI CRISPR-Cas system, we
isolated total RNA samples from E. coli cells expressing either activated
(targeting samples) or non-activated (non-targeting samples) LshCas13a
effectors. Subsequently, the RNA molecules underwent sequencing using
high-throughput techniques. Each experiment was performed in three
biological replicates. The acquired data underwent processing to eliminate
technical sequences and low-quality reads, followed by alignment to the
reference genomic sequences. Subsequently, the counts of 5' end
positions of the sequenced fragments were determined, and these counts
were compared between targeting and non-targeting samples.