ABSTRACT NUMBER - S0511

A GENOME-WIDE CRISPR/CAS9 SCREEN TO IDENTIFY NOVEL THERAPEUTIC TARGETS FOR UVEAL MELANOMA


Fan Li1,2, Vikrant Singh1, Vivienne Lu1, Jinying Chen1,3, Sandy S.C. Hung4, Joanne L. Dickinson1, Phillippa Taberlay5, Anthony L. Cook6, Alex W. Hewitt1,4,7, Guei-Sheung Liu1,3,4,7

Meeting:  2018 RANZCO


Purpose: Uveal melanoma is a potentially fatal eye cancer. A leading genetic technique “genome-wide CRISPR/Cas9 screen” was applied to systematically interrogate each gene in the genome, to identify those essential for growth and proliferation of uveal melanoma.
Method: To identify essential uveal melanoma gene pathways, clonal human uveal melanoma cell line (OCM-1) and a GeCKO (genome-wide CRISPR knockout) screening strategy were employed. OCM-
1 cells stably expressing Cas9 and genome-wide sgRNA library were generated by lentiviral-based gene transduction and cultured for at least 12 pas- sages. Stable transgene integration in pooled formats facilitates screen readout using Next-Generation Sequencing following the negative selection and Model-based Analysis of Genome-wide CRISPR- Cas9 Knockout (MAGeCK) computational tool was used to identify genes that cause cells to be depleted during selection. Moreover, bioinformatic STRING analysis inputted with the identified genes was per- formed to further explore the potential pathways that involve in the survival of the uveal melanoma. Results: A total of 81 genes were found by MAGeCK with at least 3 or more targeted sgRNA deletions during selection. Bioinformatics STRING analysis revealed that the genes targeted by these identified sgRNAs are involved in many biological pathways, including mRNA processing and splicing. Conclusion: Here, we have identified genes essen- tial for the proliferation and survival of uveal mela- noma via a forward genetic “genome-wide CRISPR/ Cas9 screen”. Our work may provide new insights into the molecular mechanisms of uveal melanoma and reveal new therapeutic target to treat this poten- tially devastating disease.