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Xiaohe Yan, Dali Li, Hongbo Cheng; Generation of a Novel Peroxidasin Knockout Mouse. Invest. Ophthalmol. Vis. Sci. 2018;59(9):325.
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Mutations in Peroxidasin (PXDN) cause severe inherited eye disorders in humans, such as congenital cataract, corneal opacity and developmental glaucoma. However, its pathophysiological mechanism remains largely unknown. As there are mutiple domains in PXDN, including Leucine-rich repeats (LRR), immunoglobulin domain, peroxidase domain and VW domain. The function of LRR domain remains unknown. To investigate the role of LRR domain during the eye development and disease, we generated a novel PXDN knockout mice that two exons of LRR domains are deleted and performed preliminary characterization.
C57BL/6J mice were used and PXDN knockout mouse was created by CRISPR-Cas9 technology using two sgRNAs targeting intron 3 and 6 of Pxdn gene, a region spanning LRR3 and LRR4 domains. Then sgRNA and Cas9 mRNA were injected into the cytoplasm of one-cell stage embryos. Injected zygotes were transferred into pseudopregnant female mice immediately after injection. The PXDN knockout mice were confirmed by PCR genotyping with subsequent sequencing. The 74aa deletion was efficiently transmitted to the F1 generation.
We generated a novel PXDN mouse line with the genetic deletion of the fragment of LRR3 and LRR4 of the LRR domain. This deletion in the two exons causes severe anterior segment dysgenesis and microphthalmia resembling the manifestations in patients with PXDN mutations. It is also similar to our previous mouse model of PXDN with a point mutation in the peroxidase domain.
We generated a novel PXDN mouse line, which resulted in the genetic deletion of two repeats of the LRR domain. We found that deficiency of LRR domain of the PXDN lead to severe anterior segment dysgenesis and microphthalmia. We will further characterize detailed phenotype and deepen our understanding of peroxidasin in eye development and disease such as congenital cataract, corneal opacity and developmental glaucoma. This animal model can also be used to test potential durg therapeutics for anterior segment dysgenesis.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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