Fig. 1: CRISPR-CasRx can effectively lower sense and antisense repeat-containing transcripts and DPRs and prevent polyGR and polyPR formation in HEK293T cells. | Nature Communications

Fig. 1: CRISPR-CasRx can effectively lower sense and antisense repeat-containing transcripts and DPRs and prevent polyGR and polyPR formation in HEK293T cells.

From: Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo

Fig. 1

a Strategy for targeting sense and antisense C9orf72 transcripts with gRNAs. Sense gRNAs target repeat-containing variants 1 and 3 but not variant 2. b Sense and antisense NLuc reporter assay designs. c Sense NLuc assays testing single plasmids expressing both CasRx and sense guides. d Antisense NLuc assays testing single plasmids expressing both CasRx and antisense guides. Data in (c, d) are shown as mean ± S.D. n = 2 biological repeats (all technical repeats shown on graph), one-way ANOVA with Dunnett’s test for post-hoc analysis. e, f Single vectors expressing a single guide (guide 10 for sense targeting or guide 17 for antisense targeting) or both guides 10 and 17 were used in our sense and antisense NLuc reporter assays. CasRx with sense and antisense targeting array can effectively reduce both e sense and f antisense C9orf72 DPR levels to a similar degree to single guide expressing plasmids indicating effective guide array maturation and multi-target engagement. All NLuc data normalised to FLuc and non-targeting (NT+NT) guide. Data in (e, f) shown as mean ± S.D. n = 3 biological repeats (all technical repeats shown on graph), one-way ANOVA with Dunnett’s test for post-hoc analysis. Source data are provided as a Source Data file.

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