Fig. 6: Screening of crRNA targeting the TTR gene in human HepG2 cells.
From: CRISPR–Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis
a, Schematic representation of the human TTR gene (ENST00000237014.8), indicating the positions of the candidate crRNAs (UT01–UT10, shown as orange pentagons), the Cas9 gRNA (NTLA-2001) and ddPCR probe sites. b, Relative quantification of genome-editing efficiency measured by ddPCR in HepG2 cells transfected with individual crRNAs (UT01–UT10) or the Cas9 gRNA (NTLA-2001) (n = 2–4 independent biological replicates (separately seeded and transfected HepG2 cell cultures)). c, Relative quantification of TTR mRNA expression levels by RT–qPCR using exon 1–2 and exon 3–4 junction probes. Several Cas3 crRNAs (UT05–UT07) resulted in reduced TTR transcript levels, with magnitudes comparable to or approaching NTLA-2001 in this assay (n = 3 independent biological replicates (separately seeded and transfected HepG2 cell cultures)). Error bars indicate the mean ± s.e.m. Statistical analysis was performed using a one-way ANOVA with Dunnett’s post hoc test versus untreated HepG2 cells (****P < 0.0001; NS, not significant).
