Fig 4 s1

Fig 4 s1

Sequenced reads are mapped against the danRer11 zebrafish genome assembly with Blat and are visualized in the UCSC genome browser (black) together with the serpine3 RefSeq annotation (blue). (A) PCRs with primers For1 and Rev2 or For1 and Rev2.2 (green) amplify the expected regions around the transcription start site on chromosome 9 in wild type (WT_Rev2, WT_Rev2.2). Injection of CRISPR guides 1, 2, and 3 (orange) results in a deletion of about 400 bp for founder individuals 7 and 12 in serpine3^cbg17. Offspring of founder 12 (394 bp deletion) was raised and further crossed. The location of a single serpine3 transcription start site is supported by annotation and activating histone marks H3K4me3 within the respective region (lower gray bar with darkness of color correlating with signal intensity). (B) PCR with primers For3 and Rev4 (green) amplifies a region around coding exon1 of serpine3. Injection of CRISPR guides 4, 5, and 6 (orange) results in a deletion of about 100 bp for founders 1, 9, 18, and 19 in serpine3^cbg18. For founder 9, this 92 bp deletion induces a frame-shift in the reading frame (bold) with three early stop codons (two shown as *, third stop located in coding exon 2) and offspring was grown. The deletion is equivalent to deletion of 31 amino acids (red) and +1 nt insertion. The amino acid encoded by a split codon is shown in blue.