|
| Status |
Public on Sep 16, 2020 |
| Title |
Transposon mutagenesis screen identifies MHC class II transactivator (CIITA) as an inducer of cellular resistance to Ebola virus and SARS Coronavirus |
| Organism |
Homo sapiens |
| Experiment type |
Other
|
| Summary |
U2OS cells were co-transfected with PiggyBac (PB) transposon plasmid pPB-SB-CMV-puro-SD3 and the transposase p-hyPBASE. The modified PiggyBac (PB) transposon, which has a constitutively active CMV promoter which can stimulate or disrupt expression of neighboring genes, depending on insertional orientation. For each library, between 107-108 cells were transfected, cultured with the addition of 2 µg/ml puromycin for one week to select cells that had incorporated the transposon, and then used for screens of virus resistance with minimal further expansion. Mutagenized cells were challenged with replication-competent recombinant EboGP-VSV at MOI of 1 or 10 and propagated for up to 3 weeks with regular media changes to select virus-resistant colonies. After virus resistant cells were selected, genomic DNA (gDNA) was isolated from 5-10 X 106 cells and transposon insertion sites were mapped using high throughput sequencing.
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| |
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| Overall design |
Eight individual libraries were created and were combined for common insertion site analysis (CIS).
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| |
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| Contributor(s) |
Sha K, Lacy-Hulbert A, Bruchez A |
| Citation(s) |
32855215 |
| |
| Submission date |
Aug 21, 2020 |
| Last update date |
Jan 03, 2024 |
| Contact name |
Stephanie Osmond |
| E-mail(s) |
sosmond@benaroyaresearch.org
|
| Organization name |
Benaroya Research Institute
|
| Street address |
1201 9th Ave
|
| City |
Seattle, |
| State/province |
WA |
| ZIP/Postal code |
98101 |
| Country |
USA |
| |
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| Platforms (2) |
| GPL15456 |
Illumina HiScanSQ (Homo sapiens) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
|
| Samples (15)
|
|
| Relations |
| BioProject |
PRJNA658479 |
| SRA |
SRP278348 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE156598_SD3-040_U.L2AB.S1.E10_trimmed.bedGraph.gz |
14.0 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_SD3-041_U.L5C5.S1.E1_trimmed.bedGraph.gz |
11.2 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_SD3-054_U.L3AB.S1.E10_trimmed.bedGraph.gz |
7.0 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_SD3-301_U.L7AB.S1.E10_trimmed.bedGraph.gz |
101.0 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_SD3-304_U.L5AB.S1.E10_trimmed.bedGraph.gz |
94.7 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_mutAB-L1_trimmed.bedGraph.gz |
62.5 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_mutKS-L1G3_trimmed.bedGraph.gz |
20.8 Kb |
(ftp)(http) |
BEDGRAPH |
| GSE156598_mutKS-L1H3_trimmed.bedGraph.gz |
24.7 Kb |
(ftp)(http) |
BEDGRAPH |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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