Reduction of UreB and CagA expression level by siRNA construct in Helicobacter pylori strain SS1.

Reduction of UreB and CagA expression level by siRNA construct in Helicobacter pylori strain SS1.
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DOI:
10.1186/s12866-023-03143-x
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发表时间:
2023-12-19
期刊:
影响因子:
4.2
通讯作者:
Alvandi A
Alvandi A
中科院分区:
生物学3区
文献类型:
--
作者:
Motamedi H;Abiri R;Salari F;Jalili C;Alvandi A

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两种重要的毒力因子--尿素酶和cagA在幽门螺杆菌(H. pylori)中发挥着重要作用。pylori)胃癌。本研究的目的是利用小干扰RNA(siRNA)研究ureB和cagA的表达水平和功能。SS 1株H. pylori是自然转化的宿主。将针对ureB和cagA基因设计的siRNA插入pGPU 6/GFP/Neo siRNA质粒载体中,以使用表型和基因型方法进行评估。然后,进行qPCR以测定ureB和cagA基因表达的抑制率。重组菌株SS 1中siRNA-ureB和siRNA-cagA的表达水平分别比天然H. pylori菌株SS 1。此外,体外siRNA-ureB的初步评价显示尿素酶活性的抑制。这些数据表明,siRNA可能是一个强大的新工具,在体外基因沉默,并为基于RNAi的抗H。幽门螺杆菌治疗我们的研究结果表明,靶向ureB和cagA基因的siRNA似乎是一个新的策略,抑制脲酶酶活性,减少炎症和定植率。
Two important virulence factors, urease and cagA, play an important role in Helicobacter pylori (H. pylori) gastric cancer. Aim of this study was to investigate the expression level and function of ureB and cagA using small interfering RNAs (siRNA). SS1 strain of H. pylori was considered as host for natural transformation. siRNA designed for ureB and cagA genes were inserted in pGPU6/GFP/Neo siRNA plasmid vector to evaluate using phenotypic and genotypic approaches. Then, qPCR was performed for determining inhibition rate of ureB and cagA gene expression. The expression levels of siRNA-ureB and siRNA-cagA in the recombinant strain SS1 were reduced by about 5000 and 1000 fold, respectively, compared to the native H. pylori strain SS1. Also, preliminary evaluation of siRNA-ureB in vitro showed inhibition of urea enzyme activity. These data suggest that siRNA may be a powerful new tool for gene silencing in vitro, and for the development of RNAi-based anti-H. pylori therapies. Our results show that targeting ureB and cagA genes with siRNA seems to be a new strategy to inhibit urease enzyme activity, reduce inflammation and colonization rate.
DOI: 10.1371/journal.pone.0030786
发表时间: 2012
期刊: PloS one
影响因子: 3.7
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影响因子: 11.1
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