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Silencing acpP gene via antisense oligonucleotide-niosome complex in clinical Pseudomonas aeruginosa isolates
Research in Microbiology ( IF 2.5 ) Pub Date : 2021-04-21 , DOI: 10.1016/j.resmic.2021.103834
Yamac Tekintas 1 , Devrim Demir-Dora 2 , Bayrı Erac 3 , Yasemin Erac 4 , Ozlem Yilmaz 5 , Sabire Sohret Aydemir 6 , Zuhtu Tanil Kocagoz 7 , Mine Hosgor-Limoncu 3
Affiliation  

Pseudomonas aeruginosa, an opportunistic Gram-negative pathogen, is one of the major causes of nosocomial infections. In addition to its physiological adaptation capacity, it can develop resistance to disinfectants and antibiotics through various mechanisms. Recently, new eradication methods are gaining attention. Therefore, in this study, an LNA-2′-O-methyl hybrid antisense oligonucleotide targeting the acyl carrier protein P (acpP) gene was introduced into P. aeruginosa isolates. The design was determined through sequence analysis and prediction of the secondary structure of mRNA by software. Niosomes were used for enhancing cellular uptake. The control of the binding and transfection ability of the sequence was determined fluorometrically by labeling with 6-Fam. The effects were determined with broth microdilution method and qPCR studies. Eight different formulations were prepared. Among these, one formulation has shown to have ASO complexation ability whose composition was 312 μl Span 80 + 69.5 mg Cholesterol+ 36.4 mg CTAB+1 ml Chloroform and 5 ml dH2O. Thus this formulation was determined as the delivery system for the next stages. Significant gene inhibition was detected at the six isolates. Results of this study suggested that niosomes can be used as a delivery system for cellular uptake of ASO and could eliminate bacterial growth.



中文翻译:

在临床铜绿假单胞菌分离物中通过反义寡核苷酸-niosome 复合物沉默 acpP 基因

铜绿假单胞菌是一种机会性革兰氏阴性病原体,是医院感染的主要原因之一。除了生理适应能力外,它还可以通过各种机制产生对消毒剂和抗生素的抗性。最近,新的根除方法越来越受到关注。因此,在本研究中,将靶向酰基载体蛋白 P ( acpP ) 基因的 LNA-2'-O-甲基杂合反义寡核苷酸引入铜绿假单胞菌分离株中。通过序列分析和二级结构预测确定设计mRNA 的软件。Niosomes 用于增强细胞摄取。序列的结合和转染能力的控制通过用 6-Fam 标记荧光测定法确定。使用肉汤微量稀释法和 qPCR 研究确定了效果。制备了八种不同的制剂。其中,一种配方显示具有 ASO 复合能力,其组成为 312 μl Span 80 + 69.5 mg 胆固醇 + 36.4 mg CTAB+1 ml 氯仿和 5 ml dH 2 O。因此,该配方被确定为下一阶段的递送系统. 在六个分离株中检测到显着的基因抑制。这项研究的结果表明,niosomes 可用作 ASO 细胞摄取的传递系统,并可以消除细菌生长。

更新日期:2021-04-21
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