title: CRISPR Cas9-guided chromatin immunoprecipitation identifies miR483 as an epigenetic modulator of IGF2 imprinting in tumors.
abstract: The normally imprinted insulin-like growth factor II (IGF2) gene is aberrantly upregulated in a variety of human malignancies, yet the mechanisms underlying this dysregulation are still poorly defined. In this report, we used a CRISPR Cas9-guided chromatin immunoprecipitation assay to characterize the molecular components that participate in the control of IGF2 gene expression in human tumor cells. We found that miR483, an oncogenic intronic miRNA, binds to the most upstream imprinted IGF2 promoter, P2. Ectopic expression of miR483 induced upregulation of IGF2 expression, in parallel with an increase in tumor cell proliferation, migration, invasion, and tumor colony formation. miR483 induced loss of IGF2 imprinting by altering the epigenotype at P2, with reduction in histone H3K27 methylation and a decrease in chromatin binding of two imprinting regulatory factors, CTCF and SUZ12. This study identifies a new role for miR483 in the regulation of IGF2 gene expression through the alteration of the promoter epigenotype.
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.|Department of Medicine, Stanford University Medical School, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA.
authors.forename: Yiqun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.|Department of Medicine, Stanford University Medical School, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA.
authors.forename: Ji-Fan
authors.initials: JF
authors.lastname: Hu
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.|Department of Medicine, Stanford University Medical School, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.
authors.forename: Jiuwei
authors.initials: J
authors.lastname: Cui
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.
authors.forename: Sujun
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Medicine, Stanford University Medical School, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Hoffman
authors.affiliation: Stem Cell and Cancer Center, First Affiliated Hospital, Jilin University, Changchun, Jilin 130061, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
mesh_terms: D064113:CRISPR-Cas Systems; D002465:Cell Movement; D049109:Cell Proliferation; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D015972:Gene Expression Regulation, Neoplastic; D018392:Genomic Imprinting; D045325:HCT116 Cells; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015854:Up-Regulation
chemical_list: C035170:IGF2 protein, human; C548912:MIRN483 microRNA, human; D035683:MicroRNAs; D007335:Insulin-Like Growth Factor II
keywords: IGF2 imprinting; allelic expression; epigenetics; histone K27 methylation; tumor