CRISPR/Cas9-mediated knockout of mmp13a, mmp13b, slc5a1, and slc5a2 in a hypoxia-induced zebrafish embryo heart failure model
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Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
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Heart failure (HF) is a complex condition where the heart is unable to provide sufficient blood supply to meet the metabolic need. Myocardial infarction (MI) is the major cause for developing heart failure, and the prevalence of heart failure is over 10% in individuals over the age of 70. The zebrafish (Danio rerio) is widely used as an animal model for cardiovascular diseases, including heart failure. The highly conserved hypoxia response in vertebrates makes zebrafish a suitable disease model to study heart failure. In this study, I aimed to evaluate the role of the mmp13a, mmp13b, slc5a1, and slc5a2 genes in heart failure in a hypoxia-induced zebrafish embryo heart failure model using CRISPR/Cas9-mediated gene knockout techniques. The target genes were knocked out by microinjecting the Cas9 ribonucleoprotein (RNP) complex into the early-stage zebrafish embryos. The embryos were exposed to hypoxia at three days post-fertilization (3 dpf), and the heart was imaged at 4 dpf. The diameter of the heart at diastole and systole was measured, and the fractional shortening (FS%) was calculated. There was no difference in FS% after gene knockout without hypoxia exposure. After 45 minutes of hypoxia exposure, a reduction in the FS% was observed in the mmp13b knockout group, and after 60 minutes of hypoxia exposure, an improvement in the FS% was observed in the slc5a1 and slc5a2 knockout groups. These results suggest that knockout of slc5a1 and slc5a2 improves cardiac function (FS%), while mmp13b knockout impairs it under hypoxia-induced heart failure conditions in zebrafish embryos.