Abstract: Based on the American Heart Association findings, in 2020 deaths due to cardiovascular diseases made up around 30.23% of all registered deaths. According to the World Health Organisation, these diseases are among the leading causes of death worldwide. Unlike human hearts, the heart of the Danio rerio species is able to regenerate throughout its life. One of the key factors for this process is a gene found in the cardiomyocytes of zebrafish called – tbx5a, which encodes a T-box transcription factor protein. This protein plays a part not only in heart regeneration, but also heart and forelimb development. It has been discovered that mutations in the orthologue of this gene, TBX5, can lead to the development of Holt-Oram syndrome in humans. This syndrome is characterised by developmental defects of the forelimbs and heart, which in some cases may be lethal. As tbx5a has a similar principle of action to TBX5, it is important for a better understanding of heart development and regeneration. Another gene Another gene affecting zebrafish heart regeneration is tcf21, which is involved in epicardial regeneration, but interest in this gene has emerged only recently, therefore its principles of action, similarly to the tbx5a gene, are not fully understood. In this study it was attempted to construct a mechanism to analyse the binding sites of the transcription factors encoded by the tbx5a and tcf21 genes in the genome of Danio rerio cardiomyocytes. The SssI methyltransferase was chosen for that purpose. The methyltransferase gene was combined with the tbx5a gene in the pCS2+cMyc Dam_f_tbx5a plasmid and the tcf21 gene in the pCS2+cMyc Dam_f_tcf21 plasmid Afterwards, the combined genes were transcribed into mRNA, which was then injected into zebrafish embryos. A dose of 300 pg of this mRNA was shown to be non-lethal to embryos and can be used in further experiments.
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