A functional screen of RNA binding proteins identifies genes that promote or limit the accumulation of CD138+ plasma cells

  1. David J Turner
  2. Alexander Saveliev
  3. Fiamma Salerno
  4. Louise S Matheson
  5. Michael Screen
  6. Hannah Lawson
  7. David Wotherspoon
  8. Kamil R Kranc
  9. Martin Turner  Is a corresponding author
  1. The Babraham Institute, United Kingdom
  2. 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.

The following data sets were generated

Article and author information

Author details

  1. David J Turner

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Alexander Saveliev

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Fiamma Salerno

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Louise S Matheson

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Michael Screen

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  6. Hannah Lawson

    Queen Mary University of London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. David Wotherspoon

    Queen Mary University of London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Kamil R Kranc

    Queen Mary University of London, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Martin Turner

    Immunology Programme, The Babraham Institute, Cambridge, United Kingdom
    For correspondence
    martin.turner@babraham.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3801-9896

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

  1. 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

  1. Received: July 19, 2021
  2. Preprint posted: July 21, 2021 (view preprint)
  3. Accepted: April 21, 2022
  4. Accepted Manuscript published: April 22, 2022 (version 1)
  5. 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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  1. David J Turner
  2. Alexander Saveliev
  3. Fiamma Salerno
  4. Louise S Matheson
  5. Michael Screen
  6. Hannah Lawson
  7. David Wotherspoon
  8. Kamil R Kranc
  9. Martin Turner
(2022)
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

Share this article

https://doi.org/10.7554/eLife.72313

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