miRWalk is an open-source platform that provides an intuitive interface for predicting and validating microRNA (miRNA)-binding sites in known genes of human, mouse, rat, dog, and cow. The core of miRWalk is the TarPmiR algorithm, which uses a random forest approach to predict miRNA target sites across the entire transcript sequence, including the 5′-UTR, CDS, and 3′-UTR. The platform integrates results from other databases, such as TargetScan and miRDB, to enhance prediction accuracy. miRWalk is designed with a modular and extensible architecture and is updated every six months. The web interface allows users to search for miRNA or gene targets, filter results, and visualize interactions using a node graph. The database is available via Python, MySQL, and HTML/Javascript, and is deployed on the heiCloud Platform. miRWalk aims to provide up-to-date and accurate information on miRNA-target interactions, facilitating research in gene regulation and disease understanding.miRWalk is an open-source platform that provides an intuitive interface for predicting and validating microRNA (miRNA)-binding sites in known genes of human, mouse, rat, dog, and cow. The core of miRWalk is the TarPmiR algorithm, which uses a random forest approach to predict miRNA target sites across the entire transcript sequence, including the 5′-UTR, CDS, and 3′-UTR. The platform integrates results from other databases, such as TargetScan and miRDB, to enhance prediction accuracy. miRWalk is designed with a modular and extensible architecture and is updated every six months. The web interface allows users to search for miRNA or gene targets, filter results, and visualize interactions using a node graph. The database is available via Python, MySQL, and HTML/Javascript, and is deployed on the heiCloud Platform. miRWalk aims to provide up-to-date and accurate information on miRNA-target interactions, facilitating research in gene regulation and disease understanding.