|
Status |
Public on Aug 13, 2020 |
Title |
Day 3 sgAAVS1 Rep. 3 |
Sample type |
SRA |
|
|
Source name |
BC-3
|
Organism |
Homo sapiens |
Characteristics |
cell type: BC-3 Cas9 cells genotype: WT
|
Treatment protocol |
Cas9 or dCas9-KRAB cells were infected with lentiviruses expressing sgRNAs against AAVS1, IRF4, BATF, MDM4 or vIRF3.
|
Growth protocol |
All cells were grown in RPMI-1640 containing 10 µg/mL gentamycin, 0.05 mM β-mercaptoethanol and 20% Serum Plus-II media. Cells were maintained at densities between 2E5 and 1E6 cells/mL
|
Extracted molecule |
polyA RNA |
Extraction protocol |
Harvested cell pellets were resuspended in 1 mL TRIzol reagent and purified using Direct-zol RNA Miniprep with in-column DNAseI digestion. Total RNA was eluted in 50 µL water. mRNA-Seq libraries were constructed by the University of Chicago Genomics Facility using the TruSeq RNA Library Prep Kit.
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|
|
Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina HiSeq 4000 |
|
|
Data processing |
mRNA-Seq reads were aligned to hg19 and KSHV genomes using STAR-Seq. Raw read counts were generated using HTSeq. Differential gene expression analysis was performed using DESeq2 (KO/KD vs. respective AAVS1 control) Genome_build: hg19, KSHV (BAC16 reference) Supplementary_files_format_and_content: FPKM or HTSeq counts
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|
|
Submission date |
Jun 13, 2019 |
Last update date |
Aug 13, 2020 |
Contact name |
Eva Gottwein |
E-mail(s) |
[email protected]
|
Organization name |
Northwestern University
|
Department |
Microbiology-Immunology
|
Street address |
320 E Superior St., Tarry building, Room 6-752
|
City |
Chicago |
State/province |
IL |
ZIP/Postal code |
60611 |
Country |
USA |
|
|
Platform ID |
GPL20301 |
Series (1) |
GSE132707 |
mRNA-Seq of BC-3 IRF4 KO, BATF KO, and vIRF3 KD |
|
Relations |
BioSample |
SAMN12052176 |
SRA |
SRX6066708 |