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Background: E–mental health (EMH) offers various possibilities for mental health care delivery, with many studies demonstrating its clinical efficacy. However, the uptake of EMH technologies by mental health care professionals remains to be low. One of the reasons for this is the lack of knowledge and skills in using these technologies. Skill enhancement by means of serious gaming has been shown to be effective in other areas but has not yet been applied to the development of EMH skills of mental health care professionals. Objective: The aim of this paper is to describe a study protocol for the user requirements analysis for the design of a game-based training environment for mental health care professionals to enhance their skills in EMH. Methods: The user requirements are formulated using three complementary outputs: personas (lively descriptions of potential users), scenarios (situations that require EMH skills), and prerequisites (required technical and organizational conditions). We collected the data using a questionnaire, co-design sessions, and interviews. The questionnaire was used to determine mental health care professionals' characteristics, attitudes, and skill levels regarding EMH and was distributed among mental health care professionals in the Netherlands. This led to a number of recognizable subuser groups as the basis for personas. Co-design sessions with mental health care professionals resulted in further specification of the personas and an identification of different user scenarios for the game-based training environment. Interviews with mental health care professionals helped to determine the preferences of mental health care professionals regarding training in EMH and the technical and organizational conditions required for the prospective game-based training environment to be used in practice. This combination of requirement elicitation methods allows for a good representation of the target population in terms of both a broad view of user needs (through the large N questionnaire) and an in-depth understanding of specific design requirements (through interviews and co-design). Results: The questionnaire was filled by 432 respondents; three co-design sessions with mental health care professionals and 17 interviews were conducted. The data have been analyzed, and a full paper on the results is expected to be submitted in the first half of 2021. Conclusions: To develop an environment that can effectively support professionals' EMH skill development, it is important to offer training possibilities that address the specific needs of mental health care professionals. The approach described in this protocol incorporates elements that enable the design of a playful training environment that is user driven and flexible and considers the technical and organizational prerequisites that influence its implementation in practice. It describes a protocol that is replicable and provides a methodology for user requirements analyses in other projects and health care areas. International Registered Report Identifier (IRRID): RR1-10.2196/18815 JMIR Res Protoc 2021;10(2):e18815 doi:10.2196/18815 We help JMIR researchers to raise funds to pursue their research and development aimed at tackling important health and technology challenges. If you would like to show your support for this author, please donate using the button below. The funds raised will directly benefit the corresponding author of this article (minus 8% admin fees). Your donations will help this author to continue publishing open access papers in JMIR journals. Donations of over $100 may also be acknowledged in future publications. Submit Suggested contribution levels: $20/$50/$100 [ABSTRACT FROM AUTHOR]