Background: Construct validity refers to the degree to which tests that claim to measure a “construct” (i.e., an inferred concept that is intangible regarding an individual’s health or internal state such as a disease, or postulated attribute) are truly reflective of that specific construct. It is suggested that construct validity is an important concept in preclinical research, as it may help reduce misinterpretations of study results allowing for better ability to predict the success of clinical translation of preclinical studies. However, there is a lack of empirical evidence to confirm its impact on preclinical research efficacy. Objectives: (I) Conduct a scoping review of the construct validity literature as it relates to the design of in vivo animal studies. (II) Conduct an overview of systematics reviews evaluating the application and reporting of construct validity within systematic reviews of in vivo animal studies. Methods: For the scoping review, we searched Embase, MEDLINE, and Google Scholar. Eligibility criteria was intentionally broad as we included any article that mentioned construct validity in preclinical in vivo research. Further review of citations was performed on eligible studies that provided substantial discussion on construct validity. For the overview, we searched MEDLINE, Embase, and TOXLINE for systematic reviews of preclinical in vivo interventions. The outcomes of interest were the prevalence of systematic reviews that mentioned construct validity and the prevalence of reviews that assessed construct validity. Results: The literature searches for the scoping and overview yielded 3657 and 2356 articles, respectively. After screening 372 and 444 met inclusion criteria for the scoping and overview. Six codes were generated (theory; mechanism; matches the human condition; measures what it reports to; experimental conditions; and outcomes) from the content analysis for the definition of construct validity. Of the 444 systematic reviews, seven mentioned construct validity, but only three used the term construct validity directly. None of the systematic reviews assessed construct validity. Discussion/Conclusion: Construct validity was not defined uniformly among studies suggesting it is not clearly understood. There was limited reporting on construct validity in systematic reviews and entirely no assessment of it; this may reflect a lack of awareness of this concept. Future research should aim to find a consensus on the definition of construct validity in order to develop tools and frameworks to help researchers assess construct validity.