Abstract
Purpose :
Knockout mice are used in a variety of ways: they allow to test the specific functions of particular genes and to survey the processes that these particular genes may regulate. In our previous study, we found that MGST2 was a potential candidate of susceptibility genes for comitant strabismus in Japanese population. The aim of this study was to establish a MGST2 gene knockout (KO) mouse model, to use the model to search for the role of MGST2 gene, and to observe any structural anomalies of the retina and brain on MGST2 knockout mouse for further analysis.
Methods :
The CRISPR/Cas9 system with Cas9 endonuclease and guide RNAs complementary to the target was sufficient for RNA-guided cleavage of the target DNA. We introduced Cas9 protein and guide RNAs into the zygotes of C57BL/6J mouse strain to make MGST2 knockout mouse by CRISPR/Cas9 technology.
Results :
Live birth rate after embryo transfer was 28.4% (19/67); however, only 3 mice survived beyond weaning stage. Two of them (66.7%) had a deletion allele in the MGST2 locus. Homozygous mice, probably lethal, were not obtained. Heterozygous mice appeared to be healthy and normal, and showed normal size and growth of eyes, and are phenotypically indistinguishable from the wild-type.
Conclusions :
The new animal model, MGST2 knockout mouse has been established. The knockout mouse model of MGST2 will facilitate investigation into the pathogenesis of strabismus and provide opportunities to evaluate for prevention of strabismus.
In the next step, we are going to use MGST2 knockout mouse to analyze structure and function of the retina and brain by immunohistochemical staining and functional magnetic resonance imaging (fMRI).
This is a 2020 ARVO Annual Meeting abstract.