Abstract
Zebrafish have emerged as an alternative vertebrate model for both basic and applied science. While two zebrafish lines, AB and TU, have been extensively used, recent studies suggest that complex behaviors and susceptibility to adult phenotypes vary among lines. Given the increasing demand across diverse research fields, establishing a phylogenetically distinct wild-type zebrafish line without deleterious genetic variants would greatly benefit the research community. In this study, we documented the establishment of the RIKEN Wild-type (RW) line and conducted comparative genome analyses to investigate the genetic characteristics of various wild-type zebrafish lines such as AB, TU, TL, WIK, SAT, NHGRI-1, PET, *AB, IND, M-AB and IM with a particular focus on the genetic characterization of the RW line. We identified numerous genetic variants in each line that may affect coding proteins, some of which are unique to each line, conferring specific genetic traits. Notably, the RW line was found to carry such genetic variants in 13 genes. Furthermore, our phylogenetic analysis revealed that the RW line is genetically distinct from other commonly used lines. Collectively, the RW line is a robust zebrafish line with excellent breeding characteristics, making it valuable for studies exploring genetic diversity and line-specific traits within the species.