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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Aug 22, 2024 |
| Title |
ERH regulates type II interferon immune signaling through post-transcriptional regulation of JAK2 mRNA [2] |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Type II interferon (IFNγ) signaling is essential for innate immunity and critical for effective immunological checkpoint blockade in cancer immunotherapy. Genetic screen identification of post-transcriptional regulators of this pathway has been challenging since such factors are often essential for cell viability. Here, we utilize our inducible CRISPR/Cas9 approach to screen for key post-transcriptional regulators of IFNγ signaling, and in this way identify ERH and the ERH-associated splicing and RNA export factors MAGOH, SRSF1, and ALYREF. Loss of these factors impairs post-transcriptional mRNA maturation of JAK2, a crucial kinase for IFNγ signaling, resulting in abrogated JAK2 protein levels and diminished IFNγ signaling. Further analysis highlights a critical role for ERH in preventing intron retention in AU-rich regions in specific transcripts, such as JAK2. This regulation is markedly different from previously described retention of GC-rich introns. Overall, these findings reveal that post-transcriptional JAK2 processing is a critical rate-limiting step for the IFNγ-driven innate immune response.
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| Overall design |
To investigate how ERH affected the expression of immune-stimulated mRNAs in murine cells, we lentivirally transduced mouse RAW264.7-iCas9 cells with sgRNA expression vectors targeting ROSA (negative control) or ERH. The targeted genes were knocked out by by five days of doxycycline-induced Cas9 expression. We then treated cells with IFNγ, IL-1beta, or MilliQ water (non-treated control) for 4 hours, after which we harvested total RNA with Trizol and performed 3’ mRNA sequencing (Quant-Seq).
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| Contributor(s) |
Soderholm A, Popitsch N |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
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| Submission date |
Aug 06, 2024 |
| Last update date |
Aug 23, 2024 |
| Contact name |
Gijs A. Versteeg |
| E-mail(s) |
gijs.versteeg@univie.ac.at
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| Organization name |
Max Perutz Labs, University of Vienna
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| Department |
Department of Microbiology
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| Lab |
Versteeg
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| Street address |
Dr. -Bohrgasse 9
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| City |
Vienna |
| ZIP/Postal code |
1030 |
| Country |
Austria |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (18)
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| GSM8442886 |
RAW264.7-dox-Cas9, sgROSA, non-treated, biol rep 1 |
| GSM8442887 |
RAW264.7-dox-Cas9, sgROSA, non-treated, biol rep 2 |
| GSM8442888 |
RAW264.7-dox-Cas9, sgROSA, non-treated, biol rep 3 |
| GSM8442889 |
RAW264.7-dox-Cas9, sgROSA, IL-1beta, biol rep 1 |
| GSM8442890 |
RAW264.7-dox-Cas9, sgROSA, IL-1beta, biol rep 2 |
| GSM8442891 |
RAW264.7-dox-Cas9, sgROSA, IL-1beta, biol rep 3 |
| GSM8442892 |
RAW264.7-dox-Cas9, sgROSA, IFNgamma, biol rep 1 |
| GSM8442893 |
RAW264.7-dox-Cas9, sgROSA, IFNgamma, biol rep 2 |
| GSM8442894 |
RAW264.7-dox-Cas9, sgROSA, IFNgamma, biol rep 3 |
| GSM8442895 |
RAW264.7-dox-Cas9, sgERH, non-treated, biol rep 1 |
| GSM8442896 |
RAW264.7-dox-Cas9, sgERH, non-treated, biol rep 2 |
| GSM8442897 |
RAW264.7-dox-Cas9, sgERH, non-treated, biol rep 3 |
| GSM8442898 |
RAW264.7-dox-Cas9, sgERH, IL-1beta, biol rep 1 |
| GSM8442899 |
RAW264.7-dox-Cas9, sgERH, IL-1beta, biol rep 2 |
| GSM8442900 |
RAW264.7-dox-Cas9, sgERH, IL-1beta, biol rep 3 |
| GSM8442901 |
RAW264.7-dox-Cas9, sgERH, IFNgamma, biol rep 1 |
| GSM8442902 |
RAW264.7-dox-Cas9, sgERH, IFNgamma, biol rep 2 |
| GSM8442903 |
RAW264.7-dox-Cas9, sgERH, IFNgamma, biol rep 3 |
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| Relations |
| BioProject |
PRJNA1144895 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE274070_RAW.tar |
5.8 Mb |
(http)(custom) |
TAR (of CSV) |
SRA Run Selector |
| Raw data are available in SRA |
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