GSE186020
Validate whether the targeting of crRNAs affected the host cell gene expression
We explored whether the targeting of crRNAs affected the host cell gene expression. To do this, we used a type LNP, termed a lipitoid, to transiently transfect human A549 lung carcinoma cells with the Cas13d mRNA and in vitro synthesized crRNA(s). At 48 hpi, we isolated cellular RNA and performed whole-cell RNA sequencing (RNA-seq). We observed high reproducibility between biological replicates. We examined the off-target effects of viral-targeting crRNA for SARS-CoV-2 or 229E as well as the NT crRNA by comparing their transcriptome profiles to that of cells expressing Cas13d alone, and we saw highly similar gene expression profiles. Furthermore, co-delivery of Cas13d and two targeting crRNAs showed a very similar transcriptome profile to that of the Cas13d alone. These data suggest that Cas13d and antiviral crRNAs are highly specific, with a minimal impact on host cell transcriptomes. Overall design: mRNA profile of A549 cells after two days Cas13d mRNA and related crRNAs transfection