title: KO_3
organism: Homo sapiens
description: Protocols: CRISPR/Cas9 mediated KO of MED1: pSpCas9(BB)-2A-GFP (PX458) was purchased from Addgene. Guided RNA (gRNA) targeting exon 5 (5'-GAAGTGCAGTTAGATCCTGCAGG-3') or exon 13 (5'-CCAACTTATGCACCCCTGTATGA-3') of MED1 was constructed into the vector. 697 cells were transient transfected by Lipofectamine 3000 (Invitrogen) and GFP+ cells were sorted on day 3 for expansion as single clones. Isolation, subcloning and sequencing of edited genomic DNA was performed from samples on day 28. Expression of MED1 was confirmed by immunoblot with anti-MED1 antibody (Bethyl laboratories, A300-793A). 697 cells were cultured in RPMI 1640 (Invitrogen) and 10% FBS. RNA was extracted with RNeasy Mini Kit (Qiagen). Libraries were generated using the TruSeq RNA Sample Preparation Kit (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-05-13T17:05:14Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-07-29T14:45:23Z
attributes.tag: External Id
attributes.value: SAMEA5817640
attributes.tag: INSDC center name
attributes.value: TMU Research Center of Cancer Translational Medicine, Taipei Medical University
attributes.tag: INSDC first public
attributes.value: 2020-05-13T17:05:14Z
attributes.tag: INSDC last update
attributes.value: 2019-07-29T14:45:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8178:KO_3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: 697
attributes.tag: cell type
attributes.value: pre-B cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: B-cell acute lymphoblastic leukemia
attributes.tag: genotype
attributes.value: CRISPR/Cas9 mediated knockout of MED1
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: sample name
attributes.value: E-MTAB-8178:KO_3
attributes.tag: scientific_name
attributes.value: Homo sapiens