The recently released U.S. Environmental Protection Agency (U.S. EPA) Supplemental Guidance for Assessing Risk from Early Life Exposure to Carcinogens (SGAC) provides guidance to account for potential increased early life susceptibility to carcinogens that are acting via a mutagenic mode of action. While determination of the mode of carcinogenic action is central to the SGAC procedures and other regulatory risk assessments, little guidance is given as to the approaches, criteria, and nature of the evidence required to define a mutagenic mode of action. The purpose of this paper is to provide a framework along with practical guidance for the process of assigning a mode of action. Strengths, weaknesses, reliability, and choice of a test battery are discussed for select bacterial, cell culture, whole animal and human cell assays. Common confounding factors of induced pathology, cytolethality, and regenerative cell proliferation in rodent cancer bioassays are discussed along with approaches to account for these effects in assigning a mode of action and in risk assessments. Specific examples are given to illustrate the complexity in generating a data set sufficient to move from the default regulatory position of assuming a genotoxic mode of action to actually assigning a nongenotoxic mode of action. A two-part framework is proposed for assigning a mode of action. First, a weight of evidence approach is used to assess mutagenic potential based on results of genetic toxicology test systems. Second, a descriptor is assigned to classify the degree to which mutagenic activity likely played a role in the mode of action of tumor formation. This option provides a more realistic way of describing the mode of action instead of being bound by the strict genotoxic vs. nongenotoxic choices.