Compound Verification: The identity, purity and concentration of compounds submitted for chemogenomic profiling is established with state-of-the art analytical chemistry methods including HPLC, LC-MS, HRMS, and NMR.
Dose-Response Curve Determination. Prior to screening, the effect of each compound of interest is assessed in a quantitative cell proliferation assay on the IRIC robotic platform. The resulting dose-response curve is used estimate the optimal screening concentration.
Chemogenomic Profiling. Genome-wide pool of CRISPR/Cas9-generated indels in the NALM-6 human pre B-cell lymphocytic cell line are cultured with compounds of interest in parallelized fashion. After sample workup, next generation sequencing (NGS) and scoring by the RANKS algorithm, relative sgRNA depletion and enrichment identifies genes whose disruption that confers sensitivity and resistance to each compound. Applications include assessment of on- and –off target effects, characterization of hits from high-throughput chemical screens, genetic effects of toxins and environmental contaminants, drug repurposing, genetically-guided structure activity relationships, prediction of drug synergism and and antagonism, and identification of novel genetic targets for drug discovery.