ZNF469 plays a critical role in regulating synthesis of ECM

Purpose: In recent years, the mutation of ZNF469 is considered to be one of the pathogenesis of brittle cornea syndrome（BCS）. We establish a znf469 mutant zebrafish line to explore its phenotype and the possible molecular mechanism. Methods: We generated the znf469 mutant zebrafish by using the CRISPR/Cas9 system. TEM is applied to examine the phenotype of the cornea in different development stage. RNA sequencing and quantitative RT-PCR are used to reveal the molecular mechanism. Results: Macroscopically, the homozygous znf469-4del zebrafish larva exhibited a curved body from 72 hpf similar to kyphoscoliosis and a noninflated swimbladder at 7 dpf. TEM reveals the extreme reduction of corneal stroma in homozygous znf469-4del zebrafish in both central and peripheral cornea since the early development stage. RNA-seq analysis demonstrates that the znf469 mutation leads to the decreased synthesis of various ECM component like collagens and proteoglycans but increased synthesis of 26S proteasome families. Conclusion: Our work indicates that znf469 is a critical gene that encoding a transcription factor, regulating the synthesis and degradation of a large number of ECM components which is also the pathologic basis of the ocular and extraocular phenotypes in the znf469 mutant zebrafish model. Comparative gene expression profiling analysis of RNA-seq data for WT and znf469-4del zebrafish larvae at 7dpf.