Profound effect of asparagine-linked glycosylation protein 11 (TgALG11) on genomic expression of Toxoplasma gondii

Toxoplasma gondii is an intracellular parasite that relies on complex molecular mechanisms to support its proliferation and survival. In this study, we investigate the role of TgALG11 in these processes and explore the underlying regulatory network. Our data demonstrate that TgALG11 is essential for maintaining T. gondii proliferation and for normal N-glycosylation. To further elucidate the molecular mechanisms underlying the essential functions of TgALG11, we performed transcriptome sequencing to identify the genes regulated by this factor. Total RNA was extracted from iTgALG11 strains cultured with 0.5 μg/ml ATc (knockdown strain) and DMSO (control), and subjected to deep sequencing using the Illumina Novaseq™ 6000 platform. The sequence reads were subsequently mapped to the T. gondii genome, resulting in a comprehensive transcriptome encompassing 8,948 genes. Among these, 934 genes were differentially expressed due to TgALG11 deficiency, including 315 downregulated and 5 upregulated genes (DESeq, |log2(FoldChange)| ≥ 1, padj ≤ 0.05). Functional enrichment analysis revealed that these genes were associated with various Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Our findings provide novel insights into the profound impact of TgALG11 on the T. gondii transcriptome, underscoring its critical role in parasite biology. Three replicates of iTgALG11 transgenic parasites cultured with 0.5 μg/ml ATc (knockdown strain) and DMSO (control) were prepared and sent to Novo gene (Beijing, China) for TgALG11-specific transcriptome construction and analysis. .

刚地弓形虫（Toxoplasma gondii）是一种依赖复杂分子机制维持增殖与存活的细胞内寄生虫。本研究旨在探讨TgALG11在该寄生虫增殖及存活过程中的功能，并解析其潜在的调控网络。本研究数据证实，TgALG11对于维持刚地弓形虫的正常增殖以及N-糖基化（N-glycosylation）过程不可或缺。为进一步阐明TgALG11核心功能的分子机制，我们通过转录组测序技术筛选该因子调控的靶基因。实验中，我们分别从经0.5 μg/ml ATc处理的iTgALG11敲低株，以及经二甲基亚砜（DMSO）处理的对照组中提取总RNA，随后使用Illumina NovaSeq™ 6000平台完成深度测序。将测序得到的读段比对至刚地弓形虫基因组后，最终得到涵盖8948个基因的完整转录组数据集。其中，934个基因因TgALG11缺失出现显著差异表达，包括315个下调基因与5个上调基因（差异分析采用DESeq软件，|log₂(折叠变化)|≥1，padj≤0.05）。功能富集分析显示，这些差异基因富集于多种基因本体（Gene Ontology，GO）条目及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路。本研究结果为理解TgALG11对刚地弓形虫转录组的深远影响提供了新视角，凸显了其在寄生虫生物学中的关键作用。我们制备了经0.5 μg/ml ATc处理的iTgALG11转基因弓形虫及DMSO处理对照组的三个生物学重复样本，并将其送至位于中国北京的诺禾致源（Novo gene）开展TgALG11特异性转录组的构建与分析工作。