Fig. 1
From: CRISPR/Cas9-mediated genome editing induces exon skipping by alternative splicing or exon deletion
sgRNA targeting Kras induces exon skipping in single cell clones. a Schematic of an sgRNA targeting exon 2 of the mouse Kras gene (sgKras). The red arrowhead denotes the Cas9 cleavage site. KP1 and KP2 cell lines were transduced with lentivirus that encodes Cas9 and sgKras. Two single-cell clones (KP1 clone and KP2 clone) harbor frameshift deletions. Black arrows indicate the positions of reverse transcription polymerase chain reaction (RT-PCR) primers. The G12D codon is underlined. b Normalized Kras read counts from RNA-sequencing (RNA-seq) analysis of KP parental cells (blue) and KP clones (red). RNA-seq was done twice for KP2 clone and three times for the other groups. “+” denotes WT allele. c RNA-seq showing partial exon 2 skipping in KP1 clones. RNA-seq numbers indicate reads spanning the indicated exon junctions. Two representative biological replicates are shown. d RT-PCR analysis of Kras mRNA detects an exon 2 skipped band. The expected band sizes are 331 bp and 209 bp. M, molecular marker. “*” denotes indels in PCR products from clones. e Scatter plot showing 22 exon events that change in both KP1 and KP2 clones. Exclusion of Kras exon 2 is the most frequent event. Ψ, Percentage Splicing Index