Using a Dual CRISPR/Cas9 Approach to Gain Insight into the Role of LRP1B in Glioblastoma

LRP1B remains one of the most altered genes in cancer, although its relevance cancer biology is still unclear. Recent advances gene editing techniques, particularly CRISPR/Cas9 systems, offer new opportunities to evaluate function large genes, such as LRP1B. Using a dual sgRNA approach, this study aimed assess impact disrupting glioblastoma cell biology. Four sgRNAs were designed for targeting two exons (1 and 85). The U87 (GB) line was transfected with PX459 vectors. To LRP1B-gene-induced alterations expression, PCR, Sanger DNA sequencing, qRT-PCR carried out. Three clones (clones B9, E6, H7) further evaluated. All presented cellular morphology, increased nuclear size, changes ploidy. Two (E6 showed significant decrease growth, both vitro vivo CAM assay. Proteomic analysis clones’ secretome identified differentially expressed proteins that had not been previously associated alterations. This demonstrates strategy can effectively edit GB cells, providing insights into deletions GBM