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. 2016 Nov 14;6:36921. doi: 10.1038/srep36921

Figure 1. Expression of Cas9 and JCPyV specific gRNA inhibited JCPyV infection.

Figure 1

(a) Depiction of the LentiCRISPR/gRNA plasmid design. EF-1a: Human elongation factor 1 alpha promoter, NLS: nuclear localization signal, hspCas9: human codon-optimized streptococcus pyogenes Cas9. (b) Experimental outline. SVG-A cells transduced with LentiCRISPR/gRNA virus with indicated gRNA were selected with puromycin and then infected with JCPyV-MAD1. (c) JCPyV specific gRNA target sites in JCPyV-MAD1 genome. (d) Immunofluorescence images showing the expression of LTAg (green) and VP1 (red) in SVG-A cells expressing Cas9 and the indicated gRNA at day 6 post-infection (highest virus inoculation dose in (e,f)). Scale bar = 100 μm (e) Quantification of the percentages of LTAg positive cells. (f) Quantification of the percentages of VP1 positive cells. Triangle represents the inoculation dose of JCPyV-MAD1 virus (5 × 107 genome copies/ml, 2.5 × 107 genome copies/ml, 1.25 × 107 genome copies/ml). **One-way ANOVA, p value < 0.01; ***one-way ANOVA, p value < 0.001. N = 3.