Introduction: The study was conducted to determine the factors affecting the depression, family burden and depression status of families of children with Down syndrome. Methods: This cross-sectional research was conducted with the mothers of 71 children with Down syndrome attending six special education and rehabilitation centers in Istanbul, Turkey that provide special education for children with Down syndrome. Data were collected using a sociodemographic information form, the Family Burden Assessment Scale for Families of Children with Intellectual Disability (FBAS) and the Beck Depression Inventory (BDI). Descriptive statistics, the Mann-Whitney U test, independent samples t-test, Kruskal Wallis test, One-way ANOVA, Pearson's correlation, and multiple regression analysis were used for the data analysis. Results: The mean age of the participating mothers was 41.59 ± 6.87 years; 93% were married and 56.3% had an education of 8 years or more. The mean BDI score in the study was 7.81 ± 6.92. According to FBAS, it was determined that 63.4% of the families felt a family burden. The mean BDI scores for mothers with a family burden were higher than among mothers without a family burden. The results of the regression analysis indicated that emotional burden and insufficiency perception were variables that significantly affected the depression status of the mothers. These variables account for 42% of the total variance for depression. Conclusions: As a result of the research, it was determined that depression scores increased as the emotional load and sense of insufficiency increased for mothers of children with Down syndrome.