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| Status |
Public on May 13, 2021 |
| Title |
Identification of LARP1 targets in naive CD4 T cells [eCLIP] |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
We investigated the role of RNMT in T cells using an Rnmt conditional knockout mouse model. We report that the mRNA cap methyltransferase, RNMT, supports naïve T cell survival and activation-induced proliferation. We demonstrate that RNMT has gene-specific impacts in T cells, selectively regulating expression of terminal polypyrimidine tract (TOP) mRNAs which are targets of the m7G-cap binding protein LARP1. These LARP1 eCLIP experiments determine the RNA binding sites of LARP1 in naive CD4 T cells from Rnmt cKO and control mice.
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| Overall design |
LARP1 seCLIP (single end cross linking immunoprecipitation- an enhanced iCLIP method) libraries were prepared from naïve CD4 T cells magnet sorted from mouse lymph nodes. Proteins were crosslinked to RNA using UV light prior to cell lysis, RNA was partially digested, then LARP1-crosslinked RNA immunoprecipitated and used to prepare libraries. Here we first compared three different antibodies for LARP1 seCLIP in CD4 T cells, then selected one antibody to prepare libraries from Rnmt fl/fl (control) and Rnmt fl/fl CD4cre+ (Rnmt cKO) CD4 T cells. Each experiment has libraries prepared with a LARP1 antibody, an isotype control antibody, and a size matched input control.
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| Contributor(s) |
Galloway A, Cowling VH |
| Citation(s) |
34125914 |
| |
| Submission date |
Oct 28, 2020 |
| Last update date |
Jun 21, 2021 |
| Contact name |
Alison Galloway |
| E-mail(s) |
a.galloway@beatson.gla.ac.uk
|
| Organization name |
Beatson CRUK
|
| Lab |
Victoria Cowling
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| Street address |
Garscube Estate, Switchback Road
|
| City |
Glasgow |
| ZIP/Postal code |
G61 1BD |
| Country |
United Kingdom |
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| Platforms (2) |
| GPL16417 |
Illumina MiSeq (Mus musculus) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (12)
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| This SubSeries is part of SuperSeries: |
| GSE160328 |
RNMT regulates the CD4 T cell transcriptome and translatome |
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| Relations |
| BioProject |
PRJNA672953 |
| SRA |
SRP289491 |
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