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首页> 美国卫生研究院文献>PLoS Neglected Tropical Diseases >Genetically Modifying the Insect Gut Microbiota to Control Chagas Disease Vectors through Systemic RNAi
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Genetically Modifying the Insect Gut Microbiota to Control Chagas Disease Vectors through Systemic RNAi

机译：基因改造昆虫肠道菌群通过系统RNAi控制南美锥虫病载体

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Technologies based on RNA interference may be used for insect control. Sustainable strategies are needed to control vectors of Chagas disease such as Rhodnius prolixus. The insect microbiota can be modified to deliver molecules to the gut. Here, Escherichia coli HT115(DE3) expressing dsRNA for the Rhodnius heme-binding protein (RHBP) and for catalase (CAT) were fed to nymphs and adult triatomine stages. RHBP is an egg protein and CAT is an antioxidant enzyme expressed in all tissues by all developmental stages. The RNA interference effect was systemic and temporal. Concentrations of E. coli HT115(DE3) above 3.35 × 107 CFU/mL produced a significant RHBP and CAT gene knockdown in nymphs and adults. RHBP expression in the fat body was reduced by 99% three days after feeding, returning to normal levels 10 days after feeding. CAT expression was reduced by 99% and 96% in the ovary and the posterior midgut, respectively, five days after ingestion. Mortality rates increased by 24-30% in first instars fed RHBP and CAT bacteria. Molting rates were reduced by 100% in first instars and 80% in third instars fed bacteria producing RHBP or CAT dsRNA. Oviposition was reduced by 43% (RHBP) and 84% (CAT). Embryogenesis was arrested in 16% (RHBP) and 20% (CAT) of laid eggs. Feeding females 105 CFU/mL of the natural symbiont, Rhodococcus rhodnii, transformed to express RHBP-specific hairpin RNA reduced RHBP expression by 89% and reduced oviposition. Modifying the insect microbiota to induce systemic RNAi in R. prolixus may result in a paratransgenic strategy for sustainable vector control.
机译：基于RNA干扰的技术可用于昆虫控制。需要可持续的策略来控制南美锥虫病的载体，例如Rhodnius prolixus。可以对昆虫微生物区系进行修饰，以将分子递送至肠道。在这里，表达双生红球菌血红素结合蛋白（RHBP）和过氧化氢酶（CAT）的dsRNA的大肠杆菌HT115（DE3）被喂食到若虫和成年三氢嘧啶阶段。 RHBP是一种卵蛋白，CAT是一种在所有发育阶段的所有组织中表达的抗氧化酶。 RNA干扰作用是全身性和暂时性的。在若虫和成虫中，大肠杆菌HT115（DE3）的浓度高于3.35×10 7 CFU / mL时，会产生显着的RHBP和CAT基因敲低。喂食后三天，脂肪体内的RHBP表达降低了99％，喂食后10天恢复到正常水平。摄取后五天，卵巢和后中肠的CAT表达分别降低了99％和96％。用RHBP和CAT细菌喂养的第一龄幼鸟的死亡率增加了24-30％。喂食产生RHBP或CAT dsRNA的细菌，第一龄幼虫的蜕皮率降低了100％，第三龄幼虫的蜕皮率降低了80％。产卵量减少了43％（RHBP）和84％（CAT）。胚胎发生在16％（RHBP）和20％（CAT）的产卵中被阻止。饲喂雌性10 5 CFU / mL的天然共生菌Rhodococcus rhodnii，使其转化为表达RHBP特异性发夹RNA，可使RHBP表达降低89％，并减少排卵。修改昆虫的微生物群以诱导螺旋藻中的系统性RNAi可能会导致对转基因策略进行可持续的载体控制。

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