Silencing of EhAQP1, EhPTP2, and EhTK Genes in Ecytonucleospora Hepatopenaei Using RNA Interference: A Potential Strategy for Preventing and Treating EHP Infection in Shrimp

IF 1.9 4区 农林科学 Q2 FISHERIES
Tingyue Liu, Yi Qiao, Huiwen Jiang, Xiaohui Cao, Jie Cheng, Ge Jiang, Xiaoman Sun, Hui Shen
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Abstract

Ecytonucleospora hepatopenaei (EHP) is an obligate intracellular parasitic pathogen of shrimp, causing growth retardation and increased susceptibility to opportunistic infections. Currently, there are few reports on the prevention and treatment of EHP infection in shrimp, and research on its drug targets is limited. This article utilizes RNA interference (RNAi) technology to conduct knockdown verification studies on potential drug targets involved in EHP’s invasion mechanism and life cycle. Three target gene sequences from genome, and annotated as EhAQP1 (aquaporin 1), EhPTP2 (polar tube protein), and EhTK (thymidine kinase) were used to synthesize specific small interfering RNA (siRNA). These siRNAs were injected into EHP-infected shrimp, and the hepatopancreas was sampled at 12, 24, 48, and 72 h postinjection to verify the interference effect and spore load. The results showed that RNAi exhibits varying inhibitory effects on different genes and is time-dependent. For EhAQP1, interference was most effective in the early stages (12 to 24 h), while the interference effects for EhTK were more pronounced at 48 and 72 h, the expression of EhPTP2 was significantly downregulated (p < 0.001) only at 24 h. In the EhAQP1 and EhPTP2 interference group, the spore load decreased from 12 to 48 h, but increased at 72 h. In contrast, the EhTK interference group showed a continuous decrease in spore load from 12 to 72 h. In summary, this study provides a new strategy for prevention and treatment of shrimp EHP infection and lays the foundation for subsequent research on RNAi–based prevention and treatment of EHP infection.

Abstract Image

利用RNA干扰沉默肝芽胞核孢子虫EhAQP1、EhPTP2和EhTK基因:一种预防和治疗虾EHP感染的潜在策略
肝芽胞核孢子虫(EHP)是虾的专性细胞内寄生病原体,可导致虾的生长迟缓和对机会性感染的易感性增加。目前,对虾EHP感染的防治报道较少,对其药物靶点的研究也有限。本文利用RNA干扰(RNAi)技术对EHP侵袭机制和生命周期中涉及的潜在药物靶点进行敲低验证研究。利用基因组中的三个靶基因序列,分别标记为EhAQP1(水通道蛋白1)、EhPTP2(极管蛋白)和EhTK(胸苷激酶),合成特异性小干扰RNA (siRNA)。将这些sirna注射到感染ehp的虾中,并在注射后12、24、48和72 h取样肝胰脏,以验证干扰效果和孢子载量。结果表明,RNAi对不同基因表现出不同的抑制作用,且具有时间依赖性。对EhAQP1的干扰在早期(12 ~ 24 h)最有效,而对EhTK的干扰在48和72 h时更为明显,EhPTP2的表达显著下调(p <;0.001),仅在24小时。在EhAQP1和EhPTP2干扰组,孢子载量从12 h下降到48 h,但在72 h增加。相比之下,EhTK干扰组在12 ~ 72 h期间孢子载量持续下降。综上所述,本研究为对虾EHP感染的防治提供了新的策略,为后续基于rnai的EHP感染防治研究奠定了基础。
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来源期刊
Aquaculture Research
Aquaculture Research 农林科学-渔业
CiteScore
4.60
自引率
5.00%
发文量
464
审稿时长
5.3 months
期刊介绍: International in perspective, Aquaculture Research is published 12 times a year and specifically addresses research and reference needs of all working and studying within the many varied areas of aquaculture. The Journal regularly publishes papers on applied or scientific research relevant to freshwater, brackish, and marine aquaculture. It covers all aquatic organisms, floristic and faunistic, related directly or indirectly to human consumption. The journal also includes review articles, short communications and technical papers. Young scientists are particularly encouraged to submit short communications based on their own research.
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