The global problem of emerging resistance of microorganisms to antibiotics makes the search for new natural substances with antibacterial properties relevant. Such substances include peptidoglycan recognition proteins (PGLYRP), which are the components of the innate immunity of many organisms, including humans. These proteins have a unique mechanism of action that allows them to evade the resistance of bacteria to them, as well as to be active against both Gram-positive and Gram-negative bacteria. However, the use of antimicrobial recombinant proteins is not always advisable due to the complexity of local delivery of the proteins and their stability; in this regard it seems appropriate to activate the components of the innate immunity. The aim of this study was to increase the expression level of native peptidoglycan recognition protein genes in HeLa cells using genome-editing technology with synergistic activation mediators (CRISPR/Cas9-SAM) and evaluate antichlamydial effect of PGLYRP. We demonstrated activation of the chlamydial two-component gene system (ctcB-ctcC), which played a key role in the mechanism of action of the peptidoglycan recognition proteins. We generated the HeLa cell line transduced with lentiviruses encoding CRISPR/Cas9-SAM activation system with increased PGLYRP gene expression. It was shown that activation of the own peptidoglycan recognition proteins gene expression in the cell line caused inhibition of the chlamydial infection development. The proposed approach makes it possible to use the capabilities of innate immunity to combat infectious diseases caused by Gram-positive and Gram-negative bacteria.

中文翻译：

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CRISPR/Cas9-SAM 介导的 HeLa 细胞中人肽聚糖识别蛋白基因表达增强抑制衣原体感染

微生物对抗生素产生抗药性的全球性问题使得寻找具有抗菌特性的新天然物质变得重要。这些物质包括肽聚糖识别蛋白 (PGLYRP)，它是包括人类在内的许多生物体先天免疫的组成部分。这些蛋白质具有独特的作用机制，使它们能够逃避细菌对其的抵抗力，并对革兰氏阳性菌和革兰氏阴性菌都具有活性。然而，由于蛋白质局部递送的复杂性及其稳定性，并不总是建议使用抗微生物重组蛋白质。在这方面，激活先天免疫的组成部分似乎是合适的。本研究的目的是使用具有协同激活介质的基因组编辑技术 (CRISPR/Cas9-SAM) 提高 HeLa 细胞中天然肽聚糖识别蛋白基因的表达水平，并评估 PGLYRP 的抗衣原体作用。我们展示了衣原体双组分基因系统 (ctcB-ctcC) 的激活，该系统在肽聚糖识别蛋白的作用机制中起关键作用。我们生成了用编码 CRISPR/Cas9-SAM 激活系统的慢病毒转导的 HeLa 细胞系，其 PGLYRP 基因表达增加。结果表明，细胞系中自身肽聚糖识别蛋白基因表达的激活导致衣原体感染发展的抑制。