title: Functional genetic screens for enhancer elements in the human genome using CRISPR-Cas9.
abstract: Systematic identification of noncoding regulatory elements has, to date, mainly relied on large-scale reporter assays that do not reproduce endogenous conditions. We present two distinct CRISPR-Cas9 genetic screens to identify and characterize functional enhancers in their native context. Our strategy is to target Cas9 to transcription factor binding sites in enhancer regions. We identified several functional enhancer elements and characterized the role of two of them in mediating p53 (TP53) and ERα (ESR1) gene regulation. Moreover, we show that a genomic CRISPR-Cas9 tiling screen can precisely map functional domains within enhancer elements. Our approach expands the utility of CRISPR-Cas9 to elucidate the functions of the noncoding genome.
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Gozde
authors.initials: G
authors.lastname: Korkmaz
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Rui
authors.initials: R
authors.lastname: Lopes
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Alejandro P
authors.initials: AP
authors.lastname: Ugalde
authors.affiliation: Division of Molecular Pathology, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Nevedomskaya
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Ruiqi
authors.initials: R
authors.lastname: Han
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Ksenia
authors.initials: K
authors.lastname: Myacheva
authors.affiliation: Division of Molecular Pathology, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Wilbert
authors.initials: W
authors.lastname: Zwart
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Ran
authors.initials: R
authors.lastname: Elkon
authors.affiliation: Division of Biological Stress Response, the Netherlands Cancer Institute, Amsterdam, the Netherlands.|Erasmus MC, Rotterdam University, Rotterdam, the Netherlands.
authors.forename: Reuven
authors.initials: R
authors.lastname: Agami
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002460:Cell Line; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D006801:Humans; D061986:MCF-7 Cells; D051379:Mice