Evaluating the Impact of gRNA SNPs in CasRx Activity for Reducing Viral RNA in HCoV-OC43

Figures

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  Figure 1. Comparing HCoV-OC43 (GenBank: AY585228) and SARS-CoV-2 (GenBank: NC_045512.2) CRISPR target sequences. Mismatches between HCoV-OC43 and SARS-CoV-2 sequences are highlighted based on nucleotide.

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  Figure 2. gRNA plasmid cloning workflow. Created in BioRender.com, accessed on 27 September 2021.

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  Figure 3. Genomic location of HCoV-OC43 CRISPR gene targets and RT-qPCR assays. CRISPR target sequence is in grey. RT-qPCR forward primers are in dark green, reverse primers are in light green, and probes are in red. (A) RdRp_ctrl CRISPR target, (B) RdRp CRISPR target, and (C) N CRISPR target.

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  Figure 4. Microscopy images of transfection efficiency 48 h post-transfection. (A) Transfected Vero cells under bright field. (B) Transfected Vero cells under GFP fluorescence.

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  Figure 5. Percent reduction of HCoV-OC43 RNA for each gRNA over time. Data presented as the mean of triplicate replicates with error bars representing the standard error.

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  Figure 6. RdRp gRNA mismatch comparison. Data presented as the mean of triplicate replicates with error bars representing the standard error. Graphs in the top row compare number of SNPs while graphs on the bottom compare the SNP location at Day 2 post-infection.

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  Figure 7. N gRNA mismatch comparison. Data presented as the mean of triplicate replicates with error bars representing the standard error. Graphs in the top row compare number of SNPs while graphs on the bottom compare the SNP location at Day 2 post-infection.