characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-01-01T17:01:42Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-24T15:23:15Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104225661
characteristics.characteristic: External Id
characteristics.text: Institute of Molecular Biology and Tumor Resarch (IMT), Philipps-University of Marburg, Marburg, Germany
characteristics.characteristic: INSDC center name
characteristics.text: 2018-01-01T17:01:42Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-24T15:23:15Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1884679
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6006:HEK_PCGF6ko_antiL3MBTL2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: HEK293
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001182
characteristics.characteristic: cell line
characteristics.text: epithelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000066
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: HEK293 cells were cultured in DMEM/F-12 + GlutaMax medium (gibco) supplemneted with 10% fetal bovine serum (Sigma) and 1 % Penicillin/Strepomycin (Sigma) Generation of MGA-, E2F6-, L3MBTL2- and PCGF6-depleted cell lines HEK293 cells were transfected using FugeneHD (Promega, City, Country) with plasmids expressing mammalian-codon optimized Cas9 and sgRNAs targeting the coding region of human MGA, E2F6, L3MBTL2 or PCGF6. The parental vector pSpCas9-2A-Puro (pX459) was a gift from Feng Zhang (Addgene plasmid # 48139) . Sequence of oligonucleotides used for targeting: MGA gRNA6: CATCTGGAAAGGTACTCCCA gRNA7: GTCATACTTGAATTGTATAC E2F6 gRNA1: GGGTATTCTTGACTTAAACA gRNA2: GTTTAAGTCAAGAATACCCC gRNA3: GTCGATTCCATCTAAGACAT L3MBTL2 gRNA6: GGATGTGATGAAAGGGATGA gRNA7: GCCTCTGTCATCCAGACAGC PCGF6 gRNA3: GGTATGAAGACATTCTGTGA gRNA4: TGTACTACTATATTGCATTT The empty pX459 vector was transfected as a negative control. Puromycin selection (3 μg/ml) was carried out 48 hours after transfection for 3 to 6 days. Individual colonies were isolated and the targeted loci were genotyped by PCR and sequenced. Cell clones with indels in the targeted locus were further analyzed by Western blotting. ChIP was performed using the One Day ChIP Kit (Diagenode) according to manufacturer's instructions. Three to four individual ChIPs were pooled and purifed on QIAquick columns (Qiagen). Five nanograms of precipitated DNA were used for indexd deep sequency library preperation using the Microplex library preparation kit v2 (Diagenode). Libraries were purified on AMPure magnetic beads (Beckman Coulter).]
characteristics.characteristic: description
characteristics.text: embryo
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: PCGF6 knockout
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: kidney
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: HEK_PCGF6ko_antiL3MBTL2
characteristics.characteristic: title