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ABSTRACT Nigeria has a high prevalence of Sickle Cell Disease (SCD) which results in significant morbidity and mortality. Premarital genetic screening for Sickle Cell Trait (SCT) is a cost-effective means of curbing the menace. This study determined the knowledge and practices of religious leaders in the Ilorin metropolis towards pre-marital genotype determination. A descriptive cross-sectional study design and multistage sampling technique were used to select 271 respondents. IBM Statistical Package for Social Sciences (SPSS) version 23 was used for data analysis. Results were presented in prose, tables, and frequency counts while the test of association between variables was done using Chi-square and Fischer's Exact test at a significance level of < 0.05 and confidence level of 95%. Less than half 93 (39.4%) of the respondents knew both parents contribute to a child acquiring SCD while up to 109 (40.2%) strongly agreed that SCD is serious and should be a hindrance to marriage. More than half of 160 (59.0%) respondents recommend pre-marital genotype counseling for intending couples and 156 (57.6%) had recommended pre-marital genotype screening for couples. Up to 165 (60.7%) of the respondents with a tertiary level of education practiced premarital screening of their congregants (p=0.004) and 127 (58.3%) of respondents with good knowledge of SCD also practiced pre-marital screening (p=0.001). Religious leaders in the study area have a considerably high level of knowledge of pre-marital genotype screening and demonstrated a moderate level of practice. Religious leaders in the study area could serve as advocates in awareness creation and community mobilization for pre-marital haemoglobin genotype screening.