Background: Due to the broad influence and reversible nature of microRNA (miRNA) on theexpression and regulation of target genes, researchers suggest that miRNAs and single nucleotide polymorphisms(SNPs) in miRNA genes interfere with 5-fluorouracil (5-FU) drug resistance in colorectalcancer chemotherapy. Methods: Computational assessment and cataloging of miRNA gene polymorphisms that target mRNAtranscripts directly or indirectly through regulation of 5-FU chemoresistance in CRC were screened outby applying various universally accessible datasets such as miRNA SNP3.0 software. Results: 1255 SNPs in 85 miRNAs affecting 5-FU resistance (retrieved from literature) were detected.Computational analysis showed that 167 from 1255 SNPs alter microRNA expression levels leading toinadequate response to 5-FU resistance in CRC. Among these 167 SNPs, 39 were located in the seedregion of 25/85 miRNA and were more critical than other SNPs. Has-miR-320a-5p with 4 SNP in seedregion was miRNA with the most number of SNPs. On the other hand, it has been identified that proteoglycanin cancer, adherents junction, ECM-receptor interaction, Hippo signaling pathway, TGF-beta signalingcascade, biosynthesis of fatty acid, and fatty acid metabolism were the most important pathwaystargeted by these 85 predicted miRNAs. Conclusion: Our data suggest 39 SNPs in the seed region of 25 miRNAs as catalog in miRNA genes thatcontrol the 5-FU resistance in CRC. These data also identify the most important pathways regulated bymiRNA.