Application of CRISPR–Cas9 technology in the treatment of chronic lymphocytic leukemia with TP53 mutations

Aim

This study proposes the implementation of clustered regularly interspaced short palindromic repeats (CRISPR)–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 effective treatment of this heterogeneous disease.

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. The gRNA consists of CRISPR RNA (crRNA) and trans-activating CRISPR RNA, (tracrRNA), sequences, responsible for target recognition and Cas9 binding, respectively. 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

Although the natural tracrRNA–crRNA mechanism operates efficiently, the use of a single RNA-guided Cas9 system is particularly attractive due to its potential for programmed DNA cleavage and genome editing. Importantly, Cas9 can bind and cleave a target sequence only if it is adjacent to a protospacer adjacent motif. 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: CRISPR–Cas9 technology represents a powerful genetic engineering tool capable of inserting, deleting, or replacing DNA.

Dr, Aurelian Udristioiu, ,MD/PhD, is certified that European Specialist in Laboratory Medicine (EuSpLM) and Member of the European Federation of Clinical Chemistry and Laboratory Medicine, EFLM Academy.

He received National and international Awards as Professional Memberships in American Society of Hematology, ASH, for  the best Project of Research in Medical Space, "Identification of the p53 Isoform Protein in Chronic Lymphoproliferative Syndromes resistant to conventional treatment by the Enzyme-linked immunosorbent assay (ELISA) system”, being finalist in Romanian Health Care Awards, on year 2023, http://aurelianudristioiu.blogspot.com