High-resolution interrogation of functional elements in the noncoding genome

• Main Authors: Fontanillas, P. (Author), Sanjana, Neville E (Contributor), Wright, Jason (Contributor), Zheng, Kaijie (Contributor), Shalem, Ophir (Contributor), Joung, Julia (Contributor), Cheng, Chia-Wei (Contributor), Regev, Aviv (Contributor), Zhang, Feng (Contributor)
• Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Biology (Contributor), Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences (Contributor), McGovern Institute for Brain Research at MIT (Contributor), Koch Institute for Integrative Cancer Research at MIT (Contributor)
• Format: Article
• Language: English
• Published: American Association for the Advancement of Science (AAAS), 2017-12-13T15:56:11Z.
• Subjects: Article
• Online Access: Get fulltext

The noncoding genome affects gene regulation and disease, yet we lack tools for rapid identification and manipulation of noncoding elements. We developed a CRISPR screen using ∼18,000 single guide RNAs targeting > 700 kilobases surrounding the genes NF1, NF2, and CUL3, which are involved in BRAF inhibitor resistance in melanoma. We find that noncoding locations that modulate drug resistance also harbor predictive hallmarks of noncoding function. With a subset of regions at the CUL3 locus, we demonstrate that engineered mutations alter transcription factor occupancy and long-range and local epigenetic environments, implicating these sites in gene regulation and chemotherapeutic resistance. Through our expansion of the potential of pooled CRISPR screens, we provide tools for genomic discovery and for elucidating biologically relevant mechanisms of gene regulation.
National Institutes of Health (U.S.) (Award F32-DK096822)
National Institute of Mental Health (U.S.) (Grant 5DP1-MH100706)
National Institute of Mental Health (U.S.) (Grant 1R01-MH110049)