WT_4
Homo sapiens
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).
ENA-FIRST-PUBLIC
2020-05-13T17:05:14Z
ENA-LAST-UPDATE
2019-07-29T14:45:23Z
External Id
SAMEA5817646
INSDC center name
TMU Research Center of Cancer Translational Medicine, Taipei Medical University
INSDC first public
2020-05-13T17:05:14Z
INSDC last update
2019-07-29T14:45:23Z
INSDC status
public
Submitter Id
E-MTAB-8178:WT_4
broker name
ArrayExpress
cell line
697
cell type
pre-B cell
common name
human
disease
B-cell acute lymphoblastic leukemia
genotype
wild type genotype
organism part
bone marrow
sample name
E-MTAB-8178:WT_4
scientific_name
Homo sapiens