In Gaucher disease (GD), the inherited deficiency of glucocerebrosidase results 
in the accumulation of glucocerebroside within lysosomes. Although almost 300 
mutations in the glucocerebrosidase gene (GBA) have been identified, the ability 
to predict phenotype from genotype is quite limited. In this study, we sought to 
examine potential GBA transcriptional regulatory elements for variants that 
contribute to phenotypic diversity. Specifically, we generated the genomic 
sequence for the orthologous genomic region ( approximately 39.4kb) encompassing 
GBA in eight non-human mammals. Computational comparisons of the resulting 
sequences, using human sequence as the reference, allowed the identification of 
multi-species conserved sequences (MCSs). Further analyses predicted the 
presence of two putative clusters of transcriptional regulatory elements 
upstream and downstream of GBA, containing five and three transcription 
factor-binding sites (TFBSs), respectively. A firefly luciferase (Fluc) reporter 
construct containing sequence flanking the GBA gene was used to test the 
functional consequences of altering these conserved sequences. The predicted 
TFBSs were individually altered by targeted mutagenesis, resulting in enhanced 
Fluc expression for one site and decreased expression for seven others sites. 
Gel-shift assays confirmed the loss of nuclear-protein binding for several of 
the mutated constructs. These identified conserved non-coding sequences flanking 
GBA could play a role in the transcriptional regulation of the gene contributing 
to the complexity underlying the phenotypic diversity seen in GD.