Application of CRISPR-Cas9 technology in the treatment of chronic lymphocytic leukemia with P53 gene mutations

Objective. This study proposes the implementation of clus­tered regularly interspaced short palindromic re­peats (CRISPR)-associated protein 9 (Cas9) technology for gene ther­apy targeting genetic mutations in human lym­pho­cytes affected by chronic lymphocytic leukemia (CLL), of­fe­ring new opportunities for the effective treatment of this hete­ro­ge­neous disease. Materials and method. CRISPR-Cas9 technology employs a specific enzyme guided by a de­signed guide RNA (gRNA) to a DNA target. The enzyme first introduces a cut at the target site and following this cleavage event, it can further disrupt the TP53 gene. Exa­mi­na­tion of the predicted secondary structure of the tracrRNA-crRNA duplex suggests that the features required for Cas9-catalyzed DNA cleavage at specific sites can be cap­tured within a single chimeric RNA. Results. Once the gRNA-Cas9 complex binds to the target DNA, Cas9 induces a double-strand break at the specified site, gene lesions, ai­ming to replace mutant TP53 genes in CLL cells through this technology. Conclusions. The gRNA plays a crucial role by directing the Cas9 protein to the DNA sequence of in­te­rest. In conclusion, CRISPR-Cas9 technology represents a powerful genetic engineering tool capable of inserting, de­le­ting or replacing DNA within an organism’s genome using these “molecular scissors”.