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| Status |
Public on Jan 09, 2024 |
| Title |
LITAF PROMOTES MEMBRANE REPAIR AGAINST PORE-FORMING PROTEINS |
| 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 10e7-10e8 cells were transfected, cultured with the addition of 3 µg/ml puromycin for one week to select cells that had incorporated the transposon, and then used for screens of resistance to alpha-toxin-induced cell death with minimal further expansion. Mutagenized cells were treated twice with alpha-toxin (500 ng/ml) for 48h to ensure the selection of only highly resistant cells. Genomic DNA (gDNA) was isolated from 5-10 X 10e6 cells and transposon insertion sites were mapped using high throughput sequencing.
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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) |
Stefani C, Lacy-Hulbert A |
| Citation(s) |
38181093 |
| |
| Submission date |
Sep 11, 2023 |
| Last update date |
Jan 10, 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) |
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| Samples (34)
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| Relations |
| BioProject |
PRJNA1015277 |
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