Gene are an important factor for the initiation of any disease which has a family history. Many genes are associated with RA other than environmental factors. The main objective of the study was to evaluate the association of genes PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175) polymorphisms in RA patients from population of Punjab, Pakistan. Blood samples of RA patients were collected from different hospitals of Sargodha. DNA was extracted, followed by PCR. Polymorphic analysis was performed in 300 rheumatoid arthritis patients and 300 healthy controls. PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175). In PADI4 gene, both homozygous mutant genotype (TT) and heterozygous (CT) of SNP rs2240340 showed signi cant association by increasing the risk of RA upto 2 folds (OR = 2.55; 95% CI = 1.57-4.15; p = 0.0002). In case of rs1748033 polymorphism, homozygous mutant genotype (TT) showed signi cant association with RA with increasing risk of disease up to 3-folds (OR = 3.46; 95% CI = 1.97-6.07; p = 0.0001), while heterozygous genotype (CT) of the same SNP showed signi cant association with RA by playing a protective role (OR = 0.57; 95% CI = 0.36-0.91; p = 0.0197). In HLA-DRB1 gene, homozygous mutant genotype (GG) of SNP rs2395175 showed no signi cant association with RA, while heterozygous genotype (AG) of the same SNP showed signi cant association with RA by playing a protective role (OR = 0.44; 95% CI = 0.27-0.71; p = 0.0009).Conclusions: Highly signi cance association of genes PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175) polymorphisms with RA was observed in Pakistani population.