The PCR check of the insertion gave no band for either flank

A TI{CRIMIC.GT14} DNA cassette has been inserted into Nep3, in a coding intron, and is predicted to gene trap all annotated transcripts of the gene. The TI{CRIMIC.GT14} cassette was inserted using the CRISPR/Cas9 technique together with a donor plasmid to drive homology directed repair. The sgRNA sequence used to target the gene was: TGAATACCCTATCTGGTGACAGG. The homology arms of the donor plasmid used were designed such that there is a small gap between the 3' end of the 5' arm and the 5' end of the 3' arm, thus the insertion of the TI{CRIMIC.GT14} cassette is predicted to be accompanied by a deletion of 18bp of genomic sequence flanking the insertion site.