Fishing for answers to hemostatic and thrombotic disease: Genome editing in zebrafish

Abstract

Over the past two decades, the teleost vertebrate Danio rerio (zebrafish) has emerged as a model for hemostasis and thrombosis. At genomic and functional levels, there is a high degree of conservation of the hemostatic system with that of mammals. Numerous features of the fish model offer unique advantages for investigating hemostasis and thrombosis. These include high fecundity, rapid and external development, optical transparency, and extensive functional homology with mammalian hemostasis and thrombosis. Zebrafish are particularly suited to genome‐wide mutagenesis experiments for the study of modifier genes. They are also amenable to whole‐organism small‐molecule screens, a feature that is exceptionally relevant to hemostasis and thrombosis. Zebrafish coagulation factor knockouts that are in utero or neonatal lethal in mammals survive into adulthood before succumbing to hemorrhage or thrombosis, enabling studies not possible in mammals. In this illustrated review, we outline how zebrafish have been employed for the study of hemostasis and thrombosis using modern genome editing techniques, coagulation assays in larvae, and in vivo evaluation of patient‐specific variants to infer causality and demonstrate pathogenicity. Zebrafish hemostasis and thrombosis models will continue to serve as a clinically directed basic research tool and powerful alternative to mammals for the development of new diagnostic markers and novel therapeutics for coagulation disorders through high‐throughput genetic and small‐molecule studies.

Loss of factor VIII in zebrafish rebalances antithrombin deficiency but has a limited bleeding diathesis

“…New developments in genome editing technology have facilitated quick and robust systems for targeted genetic modification, [35][36][37] which has paved the way for targeted mutagenesis in zebrafish to study coagulation disorders. 38 Here we report targeted ablation of zebrafish f8 and vwf using CRISPR/Cas9-mediated genome editing. We find that zebrafish possess a unique balance within their coagulation system, enabling them to withstand the loss of f8 without an overt hemorrhagic phenotype yet a modest decrease in their long-term survival, and deletion of vwf without apparent phenotype.…”

Purification and Characterization of a Novel Anticoagulant Peptide from Protein Hydrolysate of Eupolyphaga sinensis Walker

“…P9 binds to the substrate binding site on the enzyme, thus affecting the binding of the substrate to the enzyme; furthermore, the formation of enzyme-substrate-inhibitor complexes is able to affect V max and thus inhibit the enzyme activity.Recently, zebra sh has been widely used as a model animal for life science research in toxicology, genetics, and human diseases because of its advantages of rapid reproduction and high genetic homology with humans(Choi et al 2021;Qiu et al 2019). In fact, the transparency of zebra sh larvae makes it possible to observe thrombosis in zebra sh without using invasive methods, making zebra sh particularly suitable for the study of thrombosis(Raghunath et al 2022;Zhu et al 2016). In this study, we…”