Lawson Laboratory



ZFNs can be introduced into zebrafish by injecting mRNA encoding each ZFN monomer into 1-cell stage embryos. The injected ZFNs induce lesions by assembling as heterodimers at the target sequence. Upon binding, the nuclease domains heterodimerize leading to a double strand break in the spacer region between the target sites (spacer is represented in purple in the movie to the right, while the 5' and 3' sites are green and orange respectively). This break is repaired by non-homologous end joining, which can result in lesions being introduced at the target size. When coding sequence is targeted, the resulting lesion (usually a deletion or insertion) will disrupt protein function.

Re-play movie
How can ZFNs be used to make zebrafish knockouts?