Genome-Wide CRISPR-Cas9 Screen Identifies MicroRNAs That Regulate Myeloid Leukemia Cell Growth

Jared Wallace, Ruozhen Hu, Timothy L. Mosbruger, Timothy J. Dahlem, W. Zac Stephens, Dinesh S. Rao, June L. Round, Ryan M. O'Connell, Daniel T Starczynowski
2016 PLoS ONE  
Mammalian microRNA expression is dysregulated in human cancer. However, the functional relevance of many microRNAs in the context of tumor biology remains unclear. Using CRISPR-Cas9 technology, we performed a global loss-of-function screen to simultaneously test the functions of individual microRNAs and protein-coding genes during the growth of a myeloid leukemia cell line. This approach identified evolutionarily conserved human micro-RNAs that suppress or promote cell growth, revealing that
more » ... roRNAs are extensively integrated into the molecular networks that control tumor cell physiology. miR-155 was identified as a top microRNA candidate promoting cellular fitness, which we confirmed with two distinct miR-155-targeting CRISPR-Cas9 lentiviral constructs. Further, we performed anti-correlation functional profiling to predict relevant microRNA-tumor suppressor gene or microRNA-oncogene interactions in these cells. This analysis identified miR-150 targeting of p53, a connection that was experimentally validated. Taken together, our study describes a powerful genetic approach by which the function of individual microRNAs can be assessed on a global level, and its use will rapidly advance our understanding of how micro-RNAs contribute to human disease.
doi:10.1371/journal.pone.0153689 pmid:27081855 pmcid:PMC4833428 fatcat:bkvfpr73jbdkvn3ifgbkhccjj4