Every year, approximately 430,000 individuals worldwide are diagnosed with kidney cancer, some forms of which are highly metastatic. The transcription factor paired box gene 8 (PAX8) plays an important role during embryonic organ development and the development of renal cell carcinomas. This experiment aimed to assess the impact of editing the PAX8 binding site located on the hepatocyte nuclear factor 1 homeobox B (HNF1B) enhancers of clear cell renal carcinoma cells. The binding sites were edited to be non-functional using CRISPR-Cas9 and cellular fitness against cells with an edited functional binding site was evaluated. Bioinformatic analysis was conducted to examine the conservation of the paired box gene 8 across species through PhyloP scores and alignment analysis. The study confirmed the presence of the binding site using Illumina sequencing. Results indicated that cells with a non-functional paired box gene 8 binding site in the HNF1B enhancer were non-viable, while those with an edited functional binding site remained viable. Additionally, it was found that the binding sites were across species, particularly in mammals.