Background. Religious institutions found in the community not only uphold belief and cultural values but can also act as a force for positive change and development. Improved sanitation and hygiene are crucial in these institutions to decrease preventable infections due to unsanitary conditions. However, there are no studies among religious institutions on availability of latrines. Therefore, this study was conducted to assess latrine availability and associated factors among religious institutions in the Tigray Region, Ethiopia. Method. An institution-based cross-sectional study design was conducted in the Tigray Region, Northern Ethiopia. Multistage sampling was used to sample 385 religious institutions. Data were collected using a pretested, structured questionnaire and observation checklist. Logistic regression was fitted, and an odds ratio with 95% confidence interval (CI) with p value less than 0.05 was used to determine the predictors of latrine availability. Analysis was carried out using the SPSS 20TM software package. Results. In this study, latrine availability was 32.8%. It was significantly affected by currently saved money towards having a latrine (adjusted odds ratio (AOR): 0.32, 95% confidence interval (CI) [0.25, 0.42]), any messages seen, heard, or received on sanitation and hygiene (AOR: 0.43, 95% CI [0.38, 0.51]), and the place where messages were seen, heard, or received (AOR: 2.95, 95% CI [1.11, 5.55]). Conclusion. Latrine availability was very low when compared to the national target of 100% among religious institutions and was affected by the currently saved money towards having a latrine, any messages seen, heard, or received on sanitation and hygiene, and the place where the messages were received. Information regarding latrine availability should be provided to the community visiting religious institutions through available channels and promotion of practical models.