2025-04-05
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Molecular Oncology, Philipps-University Marburg
INSDC center name
public
INSDC status
ERS18928887
SRA accession
E-MTAB-14005:SKI KO-3_I_PMA
Submitter Id
ArrayExpress
broker name
MV4II
cell_line
macrophage
cell_type
not collected
collection date
human
common name
Protocols: RNA for RNA-sequencing (RNA-seq) was extracted from MV4-11 CTR and SKI KO clones treated with 1.5 ng/ml PMA or DMSO solvent control for 24 h. Generation of MV4-11 CRISPR/Cas9-mediated SKI knockout single cell clones (SKI KO) was performed as described in Feld et al. Nucleic Acids Res. 2018; 46(7) 3412–3428. A sgRNA against GFP was used as control (CTR). MV4-11 clones were cultured in RPMI (Life Technologies, Carlsbad, USA) supplemented with 10% FCS (Capricorn Scientific; Ebsdorfergrund, Germany) and 1% Penicillin/Streptomycin (P/S) (PAA Laboratories; Pasching, Austria). Selection and maintenance of lentiviral transduced cells was performed in the presence of 0.5 µg/ml puromycin. MV4-11 CTR and SKI KO clones were treated with 1.5 ng/ml PMA or DMSO solvent control for 24 h prior to harvesting. RNA was isolated using the RNeasy Mini kit (Qiagen) and RNA quality was assessed using the Experion RNA StdSens Analysis Kit (Bio‐Rad).  RNAseq libraries were prepared from total RNA with the Lexogen QuantSeq 3'-mRNA Library Prep Kit FWD for Illumina (Lexogen, 015.24) in combination with the UMI Second Strand Synthesis Module for QuantSeq FWD (Illumina, Read 1) (Lexogen, 081.96) following the manufacturer's protocol. Quality of sequencing libraries was controlled on a Bioanalyzer 2100 using the Agilent High Sensitivity DNA Kit (Agilent).
description
infant
developmental stage
acute monocytic leukemia
disease
SKI knock-out
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
SKI KO-3_I_PMA
title