2025-08-08
ENA first public
ERC000011
ENA-CHECKLIST
Nuclear Dynamics Unit UMR3664 CNRS/IC Institut Curie - Centre de Recherche
INSDC center name
public
INSDC status
ERS20948827
SRA accession
E-MTAB-14417:WT_H33SNAP_r2
Submitter Id
ArrayExpress
broker name
HeLa
cell_line
epithelial cell
cell_type
not collected
collection date
human
common name
Protocols: Cells were collected by trypsinization (1 million cells per condition). To ensure cells remained in G1/S block, trypsin and PBS used for washes were supplemented with thymidine to 2mM final. We used HeLa cells stably expressing H3.3-SNAP-HA or H3.1-SNAP-HA, and derived HIRA knockout cell lines (HIRA CRISPR/Cas9 KO) or HIRA wild-type control cells (GFP CRISPR/Cas9 KO) as described in Ray-Gallet et al. 2019. We cultured cells in DMEM complete medium with D-Glucose, L-Glutamine and Pyruvate, supplemented with 10% fetal calf serum, 100 U/mL Penicillin and 100 µg/mL Streptomycin. Cells were synchronized in G1/S via double thymidine block: subsequent exposure to 2 mM thymidine (16-18h), 24 µM 2-Deoxycytidine (6h) and 2 mM thymidine (18h). Total RNA was extracted using the RNeasy Plus Mini Kit according to the manufacturer’s instructions. The total RNA was DNase treated by RNase-free DNase (QIAGEN, 79254) using manufacturer’s instructions. We measured the quantity of isolated RNA using Nanodrop and checked the quality by Tapestation.  We used 10 ng of total RNA for library preparation using TruSeq Stranded Total RNA kit.
description
adult
developmental stage
cervical adenocarcinoma
disease
H3.3-SNAP-HA GFP CRISPR/Cas9 KO
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
uterine cervix
organism part
2
replicate
Homo sapiens
scientific_name
WT_H33SNAP_r2
title