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  1. Article ; Online: Framework for in vivo T cell screens.

    Milling, Lauren E / Markson, Samuel C / Tjokrosurjo, Qin / Derosia, Nicole M / Streeter, Ivy S L / Hickok, Grant H / Lemmen, Ashlyn M / Nguyen, Thao H / Prathima, Priyamvada / Fithian, William / Schwartz, Marc A / Hacohen, Nir / Doench, John G / LaFleur, Martin W / Sharpe, Arlene H

    The Journal of experimental medicine

    2024  Volume 221, Issue 4

    Abstract: In vivo T cell screens are a powerful tool for elucidating complex mechanisms of immunity, yet there is a lack of consensus on the screen design parameters required for robust in vivo screens: gene library size, cell transfer quantity, and number of mice. ...

    Abstract In vivo T cell screens are a powerful tool for elucidating complex mechanisms of immunity, yet there is a lack of consensus on the screen design parameters required for robust in vivo screens: gene library size, cell transfer quantity, and number of mice. Here, we describe the Framework for In vivo T cell Screens (FITS) to provide experimental and analytical guidelines to determine optimal parameters for diverse in vivo contexts. As a proof-of-concept, we used FITS to optimize the parameters for a CD8+ T cell screen in the B16-OVA tumor model. We also included unique molecular identifiers (UMIs) in our screens to (1) improve statistical power and (2) track T cell clonal dynamics for distinct gene knockouts (KOs) across multiple tissues. These findings provide an experimental and analytical framework for performing in vivo screens in immune cells and illustrate a case study for in vivo T cell screens with UMIs.
    MeSH term(s) Animals ; Mice ; CD8-Positive T-Lymphocytes ; Gene Knockout Techniques
    Language English
    Publishing date 2024-02-27
    Publishing country United States
    Document type Journal Article
    ZDB-ID 218343-2
    ISSN 1540-9538 ; 0022-1007
    ISSN (online) 1540-9538
    ISSN 0022-1007
    DOI 10.1084/jem.20230699
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  2. Article ; Online: X-CHIME enables combinatorial, inducible, lineage-specific and sequential knockout of genes in the immune system.

    LaFleur, Martin W / Lemmen, Ashlyn M / Streeter, Ivy S L / Nguyen, Thao H / Milling, Lauren E / Derosia, Nicole M / Hoffman, Zachary M / Gillis, Jacob E / Tjokrosurjo, Qin / Markson, Samuel C / Huang, Amy Y / Anekal, Praju V / Montero Llopis, Paula / Haining, W Nicholas / Doench, John G / Sharpe, Arlene H

    Nature immunology

    2023  Volume 25, Issue 1, Page(s) 178–188

    Abstract: Annotation of immunologic gene function in vivo typically requires the generation of knockout mice, which is time consuming and low throughput. We previously developed CHimeric IMmune Editing (CHIME), a CRISPR-Cas9 bone marrow delivery system for ... ...

    Abstract Annotation of immunologic gene function in vivo typically requires the generation of knockout mice, which is time consuming and low throughput. We previously developed CHimeric IMmune Editing (CHIME), a CRISPR-Cas9 bone marrow delivery system for constitutive, ubiquitous deletion of single genes. Here we describe X-CHIME, four new CHIME-based systems for modular and rapid interrogation of gene function combinatorially (C-CHIME), inducibly (I-CHIME), lineage-specifically (L-CHIME) or sequentially (S-CHIME). We use C-CHIME and S-CHIME to assess the consequences of combined deletion of Ptpn1 and Ptpn2, an embryonic lethal gene pair, in adult mice. We find that constitutive deletion of both PTPN1 and PTPN2 leads to bone marrow hypoplasia and lethality, while inducible deletion after immune development leads to enteritis and lethality. These findings demonstrate that X-CHIME can be used for rapid mechanistic evaluation of genes in distinct in vivo contexts and that PTPN1 and PTPN2 have some functional redundancy important for viability in adult mice.
    MeSH term(s) Mice ; Animals ; CRISPR-Cas Systems/genetics ; Protein Tyrosine Phosphatase, Non-Receptor Type 2/genetics ; Mice, Knockout ; Immune System ; Gene Editing
    Chemical Substances Protein Tyrosine Phosphatase, Non-Receptor Type 2 (EC 3.1.3.48) ; Ptpn2 protein, mouse (EC 3.1.3.48)
    Language English
    Publishing date 2023-11-27
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2016987-5
    ISSN 1529-2916 ; 1529-2908
    ISSN (online) 1529-2916
    ISSN 1529-2908
    DOI 10.1038/s41590-023-01689-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 5294: Show issues Location:
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    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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    ab Jg. 2022: Lesesaal (EG)
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