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High-resolution interrogation of functional elements in the noncoding genome.

ABSTRACT: 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.

SUBMITTER: Sanjana NE

PROVIDER: S-EPMC5144102 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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High-resolution interrogation of functional elements in the noncoding genome.

Sanjana Neville E NE Wright Jason J Zheng Kaijie K Shalem Ophir O Fontanillas Pierre P Joung Julia J Cheng Christine C Regev Aviv A Zhang Feng F

Science (New York, N.Y.) 20160901 6307

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 demo ...[more]

PMID: 27708104

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