Figure 3 from Engineering Oncogenic Hotspot Mutations on SF3B1 via CRISPR-Directed PRECIS Mutagenesis

CLL SF3B1 mutant cell lines recapitulates the altered splicing profile of primary CLL. A, Unsupervised clustering analysis and (B) PCA plot based on alternative splicing events of RNA sequencing data derived from primary CLL patient samples (n = 36) and cell lines with and without SF3B1 mutation. C, Density plot showing the position and frequency of cryptic 3′SS for SF3B1-mutated primary CLL samples and cell lines vs. average distance to the first AG from RefSeq reference of all canonical 3′SS. The number of cryptic 3′SS events used to calculate each density plot is indicated. D, Overlap analysis of significant splicing events associated with SF3B1 mutation (PSI > 0.1 or < −0.1 with P value < 0.05) among CLL, HG-3, MEC-1, and Nalm-6 cells. Events common to all samples or specific only to CLL and CLL cell lines are indicated. E, Stacked bar plot representation of the type of alternative splicing events found in common and CLL only splice variants. F, Heatmap showing the splicing profiles of 64 common and 33 CLL only splice variants. G, Splice variant qPCR analysis of GSAP and APBB3 alternative splicing in CLL, HG-3, MEC-1, and Nalm-6 with and without SF3B1 mutation in biological triplicates. Student unpaired t-test was used (*, P ≤ 0.05; **, P ≤ 0.01). Some figures created with BioRender.com.