A functional screen of RNA binding proteins identifies genes that promote or limit the accumulation of CD138+ plasma cells
- The Babraham Institute, United Kingdom
- Queen Mary University of London, United Kingdom
Abstract
To identify roles of RNA binding proteins (RBPs) in the differentiation or survival of antibody secreting plasma cells we performed a CRISPR/Cas9 knockout screen of 1213 mouse RBPs for their ability to affect proliferation and/or survival, and the abundance of differentiated CD138+ cells in vitro. We validated the binding partners CSDE1 and STRAP as well as the m6A binding protein YTHDF2 as promoting the accumulation of CD138+ cells in vitro. We validated the EIF3 subunits EIF3K and EIF3L and components of the CCR4-NOT complex as inhibitors of CD138+ cell accumulation in vitro. In chimeric mouse models YTHDF2-deficient plasma cells failed to accumulate.
Data availability
The sgRNA library is available upon request and from Addgene (#169082). The CRISPR/Cas9 knockout screen data and m6A-eCLIP data that support the findings of this study have been deposited in GEO with the GSE179919 accession code, and the RNA-seq data has been deposited in GEO with the GSE179281 accession code.
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The RNA m6A binding protein YTHDF2 promotes the B cell to plasma cell transitionNCBI Gene Expression Omnibus, GSE179919.
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Transcriptomic analysis of in vitro induced germinal centre-like B cells and plasmablast differentiationNCBI Gene Expression Omnibus, GSE179281.
Article and author information
Author details
Funding
Biotechnology and Biological Sciences Research Council (BBS/E/B/000C0427)
- Martin Turner
Biotechnology and Biological Sciences Research Council (BBS/E/B/000C0428)
- Martin Turner
Wellcome Trust (200823/Z/16/Z)
- Martin Turner
Biotechnology and Biological Sciences Research Council (BB/L016745/1)
- David J Turner
European Molecular Biology Organisation (ALTF 880-2018)
- Fiamma Salerno
Cancer Research UK (C29967/A26787)
- Kamil R Kranc
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Tomohiro Kurosaki, Osaka University, Japan
Ethics
Animal experimentation: All mouse experimentation was approved by the Babraham Institute Animal Welfare and Ethical Review Body and was licensed by the United Kingdom Home Office under PPL P4D4AF812.
Version history
- Received: July 19, 2021
- Preprint posted: July 21, 2021 (view preprint)
- Accepted: April 21, 2022
- Accepted Manuscript published: April 22, 2022 (version 1)
- Version of Record published: May 13, 2022 (version 2)
Copyright
© 2022, Turner et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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A functional screen of RNA binding proteins identifies genes that promote or limit the accumulation of CD138+ plasma cells
eLife 11:e72313.
https://doi.org/10.7554/eLife.72313
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