Codon usage bias and nucleotide bias are not influenced by the 5' flanking but by 3' and intronic region composition in SCID-associated genes.

Compositional constraints, selectional and mutational forces, nucleotide skews, RNA folding free energy, gene expression, protein properties, and differential expression are a few attributes that define the molecular signatures of any gene. The absence of information regarding these attributes for genes associated with severe combined immunodeficiency disorder (SCID) prompted us to take up this study. The compositional bias influenced codon bias. Overall percent T composition is the lowest among all nucleotides. However, its distribution varies markedly across different codon positions and is not lowest at all codon positions. We, for the first time, determined the influence of intergenic elements and introns on nucleotide and codon bias on genes and found that in SCID-associated genes, the 5' flanking region neither influences the nucleotide nor codon bias, contrary to the intronic and 3' flanking region, which both influence nucleotide and codon bias in SCID associated genes. Codon usage in the SCID-associated gene set significantly differs from the codon usage present in overall human codon usage for 33 out of 59 codons (excluding start, stop, and trp encoding). Analysis of differentially expressed genes revealed that out of the 10 most differentially expressed genes, 07 genes are Zn finger proteins (ZNF728, ZNF726, ZNF676, ZNF667, ZNF439, ZNF257, and ZNF208). Applying the knowledge of codon bias, rare codons, minimum free energy, and codon adaptation index, codon deoptimization was carried out, and ZNF208 was the best suitable candidate. The study opened the area for the identification of peculiar molecular features and the development of more candidates for gene therapy purposes.