MOLEKULARNE TESTY DIAGNOSTYCZNE IDENTYFIKUJĄCE NOSICIELI MUTACJI GENÓW SLC3A1 I SLC7A9 POWODUJĄCYCH CYSTYNURIĘ U KOTA DOMOWEGO – ANALIZA IN SILICO

Abstract

Cystinuria is an inherited genetic disease associated with mutations in the SLC3A1 and SLC7A9 genes. The results of studies in cats indicate heterogeneity of the disease. So far 1 mutation in the SLC3A1 gene and 5 mutations in the SLC7A9 gene have been discovered. Cystinuria is often detected too late (later in life), when adults have already had offspring. Quick and easy diagnosis of the disease is therefore important, even before the symptoms appear. The aim of this study was to design simple diagnostic tests to detect 6 mutations that can cause feline cystinuria. Bioinformatic analysis of the linkage of SLC3A1 and SLC7A9 with other proteins was performed and attempts were made to evaluate the structural and functional changes that occur due to mutations. The following programs and databases were used in the bioinformatics analysis: Ensembl, NCBI, Primer-BLAST, NEBcutter, STRING, SWISS-MODEL, Variant Effect Predictor (VEP). The proposed primers and enzymes for PCR-RFLP and AS-PCR assays enable the identification of mutation carriers. Identified proteins interactions suggest possible links between cystinuria and the development of other diseases. This may indirectly explain the heterogeneous and complex symptoms accompanying cystinuria. A mutation in exon 5 of the SLC7A9 gene (p.Asp236Asn) causes a change in the predicted spatial conformation of the protein. Using the VEP program, the effect of all mutations on protein functionality was assessed as deleterious. It is imperative that cat breeders first perform a molecular test for the mutation of the SLC7A9 gene (exon 7 at position c.881), as previous studies have shown that this is the most common mutation.