Introduction: Nonrandomized studies of interventions (NRSIs) are observational or experimental studies of the effectiveness and/or harms of interventions, in which participants are not randomized to intervention groups. There is increasingly widespread recognition that advancements in the design and analysis of NRSIs allow NRSI evidence to have a much more prominent role in decision making, and not just as ancillary evidence to randomized controlled trials (RCTs). Objective: To guide decisions about inclusion of NRSIs for addressing the effects of interventions in systematic reviews (SRs), this chapter updates the 2010 guidance on inclusion of NRSIs in Agency for Healthcare Research and Quality (AHRQ) Evidence-based Practice Center (EPC) SRs. The chapter focuses on considerations for decisions to include or exclude NRSIs in SRs. Methods: In November 2020, AHRQ convened a 20-member workgroup that comprised 13 members representing 8 of 9 AHRQ-appointed EPCs, 3 AHRQ representatives, 1 independent consultant with expertise in SRs, and 3 representatives of the AHRQ-appointed Scientific Resource Center. The workgroup received input from the full EPC Program regarding the process and specific issues through discussions at a virtual meeting and two online surveys regarding challenges with NRSI inclusion in SRs. One survey focused on current practices by EPCs regarding NRSI inclusion in ongoing and recently completed SRs. The other survey focused on the appropriateness, completeness, and usefulness of existing EPC Program methods guidance. The workgroup considered the virtual meeting and survey input when identifying aspects of the guidance that needed updating. The workgroup used an informal method for generating consensus about guidance. Disagreements were resolved through discussion. Results: We outline considerations for the inclusion of NRSIs in SRs of intervention effectiveness. We describe the strengths and limitations of RCTs, study design features and types of NRSIs, and key considerations for making decisions about inclusion of NRSIs (during the stages of topic scoping and refinement, SR team formation, protocol development, SR conduct, and SR reporting). We discuss how NRSIs may be applicable for the decisional dilemma being addressed in the SR, threats to the internal validity of NRSIs, as well as various data sources and advanced analytic methods that may be used in NRSIs. Finally, we outline an approach to incorporating NRSIs within an SR and key considerations for reporting. Conclusion: The main change from the previous guidance is the overall approach to decisions about inclusion of NRSIs in EPC SRs. Instead of recommending NRSI inclusion only if RCTs are insufficient to address the Key Question, this updated guidance handles NRSI evidence as a valuable source of information and lays out important considerations for decisions about the inclusion of NRSIs in SRs of intervention effectiveness. Different topics may require different decisions regarding NRSI inclusion. This guidance is intended to improve the utility of the final product to end-users. Inclusion of NRSIs will increase the scope, time, and resources needed to complete SRs, and NRSIs pose potential threats to validity, such as selection bias, confounding, and misclassification of interventions. Careful consideration must be given to both concerns.