From Descriptive to Functional Genomics of Leukemias Focusing on Genome Engineering Techniques

Abstract

Genome engineering makes the precise manipulation of DNA sequences possible in a cell. Therefore, it is essential for understanding gene function. Meganucleases were the start of genome engineering, and it continued with the discovery of Zinc finger nucleases (ZFNs), followed by Transcription activator-like effector nucleases (TALENs). They can generate double-strand breaks at a desired target site in the genome, and therefore can be used to knock in mutations or knock out genes in the same way. Years later, genome engineering was transformed by the discovery of clustered regularly interspaced short palindromic repeats (CRISPR). Implementation of CRISPR systems involves recognition guided by RNA and the precise cleaving of DNA molecules. This property proves its utility in epigenetics and genome engineering. CRISPR has been and is being continuously successfully used to model mutations in leukemic cell lines and control gene expression. Furthermore, it is used to identify targets and discover drugs for immune therapies. The descriptive and functional genomics of leukemias is discussed in this study, with an emphasis on genome engineering methods. The CRISPR/Cas9 system's challenges, viewpoints, limits, and solutions are also explored.

Keywords: CRISPR; DNA; genome engineering; leukemia.

Figure 1

Summary of gene editing techniques. TALEN = transcription activator-like effector nucleases, CRISPR/Cas9 = clustered regularly interspaced short palindromic repeats- associated protein 9, HR = Homologous recombination, NHEJ = Non-homologous end joining [8,24,41,42,43].

Figure 2

Applications of CRISPR/Cas9 technology in human therapy and hematology research [68]. CRISPR/Cas9 = clustered regularly interspaced short palindromic repeats- associated protein 9, HIV = Human Immunodeficiency Virus, CAR = Chimeric antigen receptor, CHIP = clonal hematopoiesis with an indeterminate potential.