Abstract: Objective To explore the functional activity of rs6858698 and its effect on susceptibility to chronic lymphocytic leukemia Methods Luciferase reporter assay was used to detect the regulatory activity of rs6858698-containing fragment. CRISPR/Cas9 system was employed to construct stable cell line with point mutation at rs6858698 locus. RNA-seq was used to identify differentially expressed genes between wildtype cells and mutant cells. Real-time quantitative PCR (RT-qPCR) and CRISPR interference (CRISPRi) assays were used to validate the results of RNA-seq. Results The rs6858698 -containing region with allele C showed significantly higher enhancer activity than the region containing allele G. A total of 140 differentially expressed genes were identified in HEK293T cells after changing the allele from G to C, These genes are mainly enriched in nine biological processes, including epigenetic modification. Conclusions rs6858698 has regulatory activity, which can affect the individual susceptibility to chronic lymphocytic leukemia by influencing the expression of genes related to the epigenetic modification.

Key words: rs6858698, CRISPR, RNA-seq, chronic lymphocytic leukemia