CRISPR-based approach to studying the role of lncRNA in cancer and cell differentiation

For most of the last 60 years of molecular biology, geneticists focused on studying only 2% of the human genome, the part that encodes for the protein coding genes. Unfortunately, despite the substantial effort, we have failed to explain critical biological questions and find cures to many life-threatening genetic disorders. However, recent years have witnessed the discovery of thousands of new genes, which were “hidden” within the 98% of the genome previously called “junk DNA” but now termed “noncoding genome.” The subgroup of long noncoding RNAs (lncRNAs) share many features with protein coding genes, but they do not have an open reading frame for protein synthesis. The mammalian genome encodes for thousands of lncRNAs, but why do we have these genes? Do these transcripts play an essential role in cell function? Furthermore, what is the role of lncRNAs in disease, and how can we exploit it for the benefit of human health? Scientists have unsuccessfully attempted to comprehensively address such questions. Recently, we established CRISPR-based gene regulation strategies for gene activation (CRISPRa) and inhibition (CRISPRi) for genome-scale forward genetic screening of lncRNAs. Using this platform, we identified novel genes involved in various biological pathways—including cell proliferation, genome integrity, stress response, apoptosis, and protein stability. We believe that using our approach, we will uncover novel features of this enigmatic group of genes, and the results may lead to new therapeutic opportunities for targeting cancer and other diseases.