Fig. 5: CRISPR-CasRx simultaneously reduces sense and antisense repeat RNAs in C9orf72 BAC transgenic mice. | Nature Communications

Fig. 5: CRISPR-CasRx simultaneously reduces sense and antisense repeat RNAs in C9orf72 BAC transgenic mice.

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

Fig. 5

a Diagram of experimental paradigm. CasRx PHP.eB AAV (8E+9 vg per animal) was injected via intracerebroventricular (ICV) injection into postnatal day 0 (P0) C9orf72 BAC mice. Tissue was harvested 8.5 months post-injection. CAG-CMV enhancer/chicken β-actin promoter, NLS nuclear localisation signal. Created in BioRender. Cammack, A. (2024) https://BioRender.com/u85i634. b C9orf72 promoter methylation assay of prefrontal cortical DNA extracted from C9orf72 BAC mice, demonstrating no overt transgene silencing within the cohort. c–e RT-qPCRs for C9orf72 transcripts in C9orf72 BAC mice treated with CRISPR-CasRx + gRNA AAVs in bulk prefrontal cortex 8.5 months post-injection. RT-qPCR for c sense repeat-containing C9orf72 transcripts (variants 1 and 3). d sense variant 2 C9orf72 transcripts, or e antisense repeat-containing C9orf72 transcripts. RT-qPCR data presented as fold change compared to non-targeting (NT+NT) gRNA CasRx PHP.eB AAV-treated mice. For (c–e), n = 13 Guide 10+17; n = 11 Guide NT+NT. e, f Levels of f polyGA and g polyGP DPRs in bulk cortical tissue from 8.5 month old animals treated with CasRx PHP.eB AAV. For (e, f), n = 14 Guide 10+17; n = 12 Guide NT+NT. Data in (c–g) are shown as mean ± SD, two-sided unpaired t-test. Source data are provided as a Source Data file.

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