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| Status |
Public on Feb 28, 2018 |
| Title |
Genome-Wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication |
| Organism |
Homo sapiens |
| Experiment type |
Other
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| Summary |
The emergence of influenza A viruses (IAV) from zoonotic reservoirs poses a great threat to human health. As seasonal vaccines are ineffective against zoonotic strains, and newly transmitted viruses can quickly acquire drug resistance, there remains a need for host- directed therapeutics against IAV. Here, we performed a genome-scale CRISPR/Cas9 knockout screen in human lung epithelial cells with a human isolate of an avian H5N1 strain. Several genes involved in sialic acid biosynthesis and related glycosylation pathways were highly enriched post-H5N1 selection, including SLC35A1, a sialic acid transporter essential for IAV receptor expression and thus viral entry. Importantly, we have identified capicua (CIC) as a negative regulator of cell intrinsic immunity, as loss of CIC resulted in heightened antiviral responses and restricted replication of multiple viruses. Therefore, our study demonstrates that the CRISPR/Cas9 system can be utilized for the discovery of host factors critical for the replication of intracellular pathogens.
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| Overall design |
A pooled library of A549 cells lacking individual genes was generated using CRISPR/Cas9 gene deletion technique (A549-GeCKO library; genome-wide CRISPR knockout library). The single guide RNA (sgRNA) representation in the A549-GeCKO library was sequenced. The A549-GeCKO library was subjected to 5 consecutive rounds of infection with a human isolate of low pathogenic H5N1 influenza virus and the sgRNA enriched in the resistant population were sequenced (Prelim Rd 5). In addition, a sequential screen was performed in biological duplicates and the sgRNA eriched after each round of infection were sequenced (Rd 1-5, Rep1-2 )
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| Contributor(s) |
Han J, Perez JT, Chen C, Li Y, Benitez A, Kandasamy M, Lee Y, Andrade J, tenOever B, Manicassamy B |
| Citation(s) |
29642015 |
| Submission date |
Feb 27, 2018 |
| Last update date |
Mar 26, 2019 |
| Contact name |
Yan Li |
| E-mail(s) |
yli22@bsd.uchicago.edu
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| Phone |
7738344316
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| Organization name |
The University of Chicago
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| Department |
Center for Research Informatics
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| Street address |
900 East 57th St.
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| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60637 |
| Country |
USA |
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| Platforms (2) |
| GPL15520 |
Illumina MiSeq (Homo sapiens) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA436088 |
| SRA |
SRP133553 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE111166_RAW.tar |
11.2 Mb |
(http)(custom) |
TAR (of CSV, TXT, XLSX) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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