CREATION OF ISOGENIC CELL LINE MODELS OF 17P DELETION TO STUDY CLONAL EVOLUTION AND MECHANISMS OF THERAPY RESISTANCE IN CLL AND DLBCL

Introduction: A deletion of chromosome 17p (del17p), on which the tumour suppressor gene TP53 is located, a prevalent genetic aberration in many cancer types. In chronic lymphocytic leukaemia (CLL), diffuse large B-cell lymphoma (DLBCL), and other B cell malignancies this chromosomal mutation associated with high fitness advantage setting therapy, most adverse clinical outcome. Whilst resistance to chemoimmunotherapies often used treatment CLL DLBCL can be explained by functional loss p53, such deactivation does not explain increase aggressive behaviour malignant cells. To study this, we aimed develop in-vitro models creating (15.6 kbp) (20.5 Mbp) deletions means CRISPR-Cas9 genome editing. Methods: The efficiency editing machineries were firstly assessed adherent human colorectal line HCT116 prior their application lines, are less amenable manipulation. removal was performed transient transfection vector encoding Cas9 as well single guide (sg) RNAs targeting upstream downstream coding region. For sgRNAs exon 2 GEMIN4 (telomeric) intron 5 MAP2K3 (centromeric) designed. PCR confirm deletions. Results: Isogenic lines hemizygous homozygous deletion, obtained (Figure 1). HG-3 cells, line, successfully transduced inducible-Cas9, subsequently for 17p. Keyword: Tumor Biology Heterogeneity No conflicts interests pertinent abstract.