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. 2022 Jan 19;8(3):eabk2485. doi: 10.1126/sciadv.abk2485

Fig. 3. RfxCas13d can target hHTT.

Fig. 3.

(A) (Top) HTT reporter and crRNA binding sites (red bars). (Bottom) crRNA target sequences. (B) CFP MFI in HEK293T cells transfected with reporter, RfxCas13d, and crRNAs. Data are normalized to cells transfected with RfxCas13d and NTG crRNA (n = 3). (C) (Top) Fluorescence image of HEK293T cells transfected with reporter, RfxCas13d, and HTT crRNA 1. (Bottom) Percentage of cells with HTT-CFP aggregates (n = 3). (B and C) “−” indicates cells transfected with reporter only. (D) HTT-CFP protein in HEK293T cells transfected with reporter, RfxCas13d, and crRNAs. CFP protein normalized to β-actin protein. (E and F) Relative HTT mRNA in HEK293T cells transfected with RfxCas13d/dRfxCas13d and crRNAs. Data are normalized to relative HTT mRNA in (E) cells transfected with RfxCas13d and NTG crRNA or (F) untreated cells, both indicated by dashed lines (n = 3). (G) Volcano plot of RNA-seq data comparing HEK293T cells transfected with RfxCas13d and HTT crRNA to untreated cells (n = 3). Red circle denotes HTT. (H and I) Number of differentially expressed genes [>1.25-fold change (FC), FDR-adjusted P < 0.01] in cells transfected with (H) RfxCas13d and crRNAs or (I) shRNAs, as compared to nontransfected cells (n = 3). All analyses are conducted 72 hours after transfection. Error bars indicate SEM. *P < 0.05; ***P < 0.001. (B to F) One-way ANOVA.