宿主遗传学强调 IFN-γ 依赖性的弓形虫基因，这些基因编码在体内 CRISPR 筛选中的分泌和非分泌毒力因子。

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

机构信息

Department of Immunoparasitology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan; Laboratory of Immunoparasitology, WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka, Japan; Department of Immunoparasitology, Center for Infectious Disease Education and Research, Osaka University, Suita, Osaka 565-0871, Japan.

出版信息

Cell Rep. 2023 Jun 27;42(6):112592. doi: 10.1016/j.celrep.2023.112592. Epub 2023 Jun 1.

DOI:10.1016/j.celrep.2023.112592

PMID:37269286

Abstract

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.

摘要

我们开发了一种体内 CRISPR 筛选系统，不仅可以富集毒力型弓形虫感染 C57BL/6 小鼠中的分泌型，还可以富集非分泌型毒力因子。值得注意的是，联合使用免疫缺陷型 Ifngr1 小鼠，突出了编码各种非分泌型蛋白的基因以及 ROP5、ROP18、GRA12 和 GRA45 等众所周知的效应子作为干扰素-γ（IFN-γ）依赖性毒力基因。筛选结果表明 GRA72 对于正常 GRA17/GRA23 定位以及与 UFMylation 相关基因的 IFN-γ依赖性作用具有重要意义。总的来说，我们的研究表明，宿主遗传学可以补充体内 CRISPR 筛选，以突出编码 IFN-γ依赖性分泌型和非分泌型毒力因子的基因。