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RESEARCH SERVICES Haploid Genetic Screens in Human Cells Unique experimental platform to identify resistance mutations for cancer therapy Haploid human cells are perfectly suited for genetic studies because the inactivation of one gene copy is sufficient to elucidate its loss-of-function phenotype. The obtained results are precise, robust and translate well into other model systems. Use Horizon’s haploid-based screening platform to elucidate the mechanism of resistance of anti-cancer drugs, and use the knowledge of such resistance genes to stratify patients and streamline clinical trials. t +43 1 9165522 140 f +43 1 9165522 16 [email protected] w www.horizon-genomics.com Horizon Genomics GmbH, Campus Vienna Biocenter 3, 1030 Vienna, Austria How do haploid genetic screens work? 1. Haploid cells are mutagenized using retroviral gene trapping. This generates a population of cells in which every cell line carries at least one gene trap insertions, thus resembling a gene knockout. 2. Mutant pools are treated with a drug or drug candidate that affects cell viability or cell proliferation. 3. Vast majority of mutants succumb to cell death, but a few drug resistant clones grow out. 4. Location of gene trap insertions is determined by amplifying adjacent sequences by PCR, and next generation sequencing. Retroviral Gene Trapping Haploid Cells Compound Treatment NGS Gene Trap Mutants Resistant Cells Sequence The ABT-737 screen in KBM-7 cells mutagenized with gene trapping The screen resulted in two significant hits: 140 • BAX: 51 independent insertions (p-value 10-111) 120 • NOXA: 39 independent insertions (p-value 10-70) Thus, results suggest important determinants for ABT-737 sensitivity and provide insights to the mechanism of action of the drug. 100 -log (p-value) Genetic inactivation of pro-apoptotic BAX or NOXA prevents induction of apoptosis by ABT-737 and makes cells resistant to the drug. BAX (51 insertions) NOXA (39 insertions) 80 60 40 20 0 0 100 200 300 400 500 600 700 800 Genes identified in screen (in alphabetical order) Why to choose genetic screens in haploid cells? UNBIASED No previous assumption on mechanism of action COMPREHENSIVE Whole genome coverage, multiple gene trap insertions per gene ROBUST Unprecedented signal / noise ratio PROVEN Many high impact publications highlight potential © Haploid Genetic Screens in Human Cells 2015 v-01 t + 44 (0)1223 655 580 (UK) or +1 (855) 772-4252 (USA) f + 44 (0)1223 655 581 [email protected] w www.horizondiscovery.com Horizon Discovery, 7100 Cambridge Research Park, Waterbeach, Cambridge, CB25 9TL, United Kingdom