Human DNA polymerase iota (Pol iota) is a Y-family DNA polymerase with unusual biochemical properties and not fully understood functions. Pol iota preferentially incorporates dGTP opposite template thymine. This property can be used to monitor Pol iota activity in the presence of other DNA polymerases, e.g. in cell extracts of tissues and tumors. We have now confirmed the specificity and sensitivity of the method of Pol iota activity detection in cell extracts using an animal model of loach Misgurnus fossilis embryos transiently expressing human Pol iota. The overexpression of Pol iota was shown to be accompanied by an increase in abnormalities in development and the frequency of pycnotic nuclei in fish embryos. Further analysis of fish embryos with constitutive or regulated Pol iota expression may provide insights into Pol iota functions in vertebrate animals.