92 research outputs found

    CRISPR screens identify genes essential for in vivo virulence among proteins of hyperLOPIT-unassigned subcellular localization in Toxoplasma

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    Tachibana Yuta, Sasai Miwa, Yamamoto Masahiro, et al. CRISPR screens identify genes essential for in vivo virulence among proteins of hyperLOPIT-unassigned subcellular localization in Toxoplasma. mBio , e01728-24 (2024); https://doi.org/10.1128/mbio.01728-24.The research field to identify and characterize genes essential for in vivo virulence in Toxoplasma gondii has been dramatically advanced by a series of in vivo clustered regularly interspaced short palindromic repeats (CRISPR) screens. Although subcellular localizations of thousands of proteins were predicted by the spatial proteomic method called hyperLOPIT, those of more than 1,000 proteins remained unassigned, and their essentiality in virulence was also unknown. In this study, we generated two small-scale gRNA libraries targeting approximately 600 hyperLOPIT-unassigned proteins and performed in vivo CRISPR screens. As a result, we identified several genes essential for in vivo virulence that were previously unreported. We further characterized two candidates, TgGTPase and TgRimM, which are localized in the cytoplasm and the apicoplast, respectively. Both genes are essential for parasite virulence and widely conserved in the phylum Apicomplexa. Collectively, our current study provides a resource for estimating the in vivo essentiality of Toxoplasma proteins with previously unknown localizations

    Host genetics highlights IFN-γ-dependent Toxoplasma genes encoding secreted and non-secreted virulence factors in in vivo CRISPR screens

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    Secreted virulence factors of Toxoplasma to survive in immune-competent hosts have been extensively explored by classical genetics and in vivo CRISPR screen methods, whereas their requirements in immune-deficient hosts are incompletely understood. Those of non-secreted virulence factors are further enigmatic. Here we develop an in vivo CRISPR screen system to enrich not only secreted but also non-secreted virulence factors in virulent Toxoplasma-infected C57BL/6 mice. Notably, combined usage of immune-deficient Ifngr1−/− mice highlights genes encoding various non-secreted proteins as well as well-known effectors such as ROP5, ROP18, GRA12, and GRA45 as interferon-γ (IFN-γ)-dependent virulence genes. The screen results suggest a role of GRA72 for normal GRA17/GRA23 localization and the IFN-γ-dependent role of UFMylation-related genes. Collectively, our study demonstrates that host genetics can complement in vivo CRISPR screens to highlight genes encoding IFN-γ-dependent secreted and non-secreted virulence factors in Toxoplasma.Tachibana Y., Hashizaki E., Sasai M., et al. Host genetics highlights IFN-γ-dependent Toxoplasma genes encoding secreted and non-secreted virulence factors in in vivo CRISPR screens. Cell Reports 42, 112592 (2023); https://doi.org/10.1016/j.celrep.2023.112592

    Quantum Phase Transitions of the Distorted Diamond Spin Chain

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    The frustrated quantum spin system on the distorted diamond chain lattice suitable for the alumoklyuchevskite is investigated using the numerical diagonalization of finite-size clusters and the level spectroscopy analysis. It is found that this model exhibits three quantum phases; the ferrimagnetic phase, the spin gap one, and the gapless Tomonaga-Luttinger liquid depending on the exchange coupling parameters. The ground state phase diagram is presented.Comment: to be published in JPS Conf. Se

    Jokoji and the Faith in Jizo : Based on the Analysis of "Jokoji-engi"

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    Magnetization Plateau of the Distorted Diamond Spin Chain

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    The frustrated quantum spin system on the distorted diamond chain lattice is investigated using the numerical diagonalization of finite-size clusters and the level spectroscopy analysis. In the previous work this system was revealed to exhibit the 1/3 magnetization plateau due to two different mechanisms depending on the coupling parameters, and the phase diagram at the 1/3 magnetization was obtained. In the present work it is found that the 1/3 magnetization plateau vanishes for sufficiently large XYXY-like coupling anisotropy. The phase diagram based on the level spectroscopy analysis is also presented.Comment: to be published in JPS Conference Proceeding
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