(A) Schematic representation of the allele-specific editing strategy. An sgRNA spanning the 4 bp insertion of NPM1 mutant A (sgNPM1c) was designed to specifically target the mutant allele.
(B) The nucleotide sequence, C-terminal amino-acid sequence, and corresponding allele frequency (n=3; mean ± SEM) of the most represented mutant alleles observed following amplicon sequencing of OCI-AML3 cells transfected with sgNPM1c.
(C) Immunofluorescence with an antibody against the N-terminus of NPM1 (total NPM1) 4 days after transfection with the indicated sgRNA. An unedited cell with cytoplasmic protein is shown (white arrow). Scale bar: 20 μm.
(D) Immunoblot of total NPM1 (top) and NPM1c C-terminal epitope (middle) 4 days after transfection with the indicated sgRNA. See also Figure S1 and Table S1.