Utilizing engineered nuclease genome editing technologies, Cyagen has generated an Immunoglobulin M (IgM) knockout rat model. IgM exon 2 was chosen as the engineered nuclease target site (Fig 1). Two pairs of nucleases were designed to cut two different sites.
Figure 1. Engineered Nuclease Knockout Strategy.
DNA sequencing result that shows the frameshift mutation in engineered nuclease knockout rats (Fig 2): Of the three rat pups obtained with the IgM gene modified, two pups had frameshift mutations in one allele (as indicated by the distorted peaks in MT rats). These founders may be subsequently bred to obtain a homozygous IgM knockout line.
Figure 2. DNA sequencing profile of knockout mutant rats (MT) compared to wild-type rats (WT).