Due to the lack of efficacy of the currently used chemical drugs, poor tick control, and lack of effective vaccines against Babesia, novel control strategies are urgently needed. In this regard, searching for anti-Babesia gene therapy may facilitate the control of this infection. Following this pattern, small interfering RNAs (siRNAs) are widely used to study gene function and hence open the way to control the parasite. However, the primary constraint of this approach is the lack of Babesia to RNA-induced silencing complex (RISC) enzymes, making siRNA impractical. In this study, we preassembled complexes with the human enzyme argonaute 2 (hAgo2) and a small interfering RNA (siRNA)/single-stranded RNA (ssRNA) against B. gibsoni and B. microti metabolite transporters. The assembled complexes were generated by developing a gene delivery system with chitosan dehydroascorbic acid nanoparticles.The delivery system effectively protected the loaded RNAi and targeted Babesia-infected RBCs with a relatively high internalization rate. The assembled complexes were successfully transfected into live parasites for specific slicing of Babesia targets. We demonstrated a reduction in the expression of target genes at the mRNA level. Furthermore, this silencing inhibited Babesia growth in vitro and in vivo. For the first time, we used this method to confirm the role of the assembled complexes in manipulating the noncanonical pathway of RNAi in Babesia parasites. This novel method provides a means of silencing Babesia genes to study their role in host-parasite interactions and as potential targets for gene therapy and control.

中文翻译：

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纳米颗粒递送的 hAgo2 和 ssRNA 的预组装复合物：一种新的沉默基因表达方法，克服了顶复门寄生虫田鼠巴贝虫（具有兽医和人畜共患学重要性的血液寄生虫）中 siRNA 加工经典途径的缺失。


由于目前使用的化学药物缺乏疗效、蜱虫控制不佳以及缺乏针对巴贝虫的有效疫苗，迫切需要新的控制策略。在这方面，寻找抗巴贝虫基因疗法可能有助于控制这种感染。按照这种模式，小干扰 RNA （siRNA） 被广泛用于研究基因功能，从而为控制寄生虫开辟了道路。然而，这种方法的主要限制是缺乏 RNA 诱导的沉默复合物 （RISC） 酶的巴贝虫，这使得 siRNA 不切实际。在这项研究中，我们与人酶 argonaute 2 （hAgo2） 和针对 B. gibsoni 和田鼠 B. 代谢物转运蛋白的小干扰 RNA （siRNA） /单链 RNA （ssRNA） 预组装复合物。组装的复合物是通过开发壳聚糖脱氢抗坏血酸纳米颗粒的基因递送系统产生的。该递送系统有效地保护了负载的 RNAi 并以相对较高的内化率靶向巴贝虫感染的红细胞。将组装的复合物成功转染到活寄生虫中，用于巴贝虫靶标的特异性切片。我们证明了靶基因在 mRNA 水平上的表达降低。此外，这种沉默抑制了巴贝虫在体外和体内的生长。我们首次使用这种方法来确认组装的复合物在巴贝虫寄生虫中操纵 RNAi 非经典途径中的作用。这种新方法提供了一种沉默巴贝虫基因的方法，以研究它们在宿主-寄生虫相互作用中的作用，并作为基因治疗和控制的潜在靶标。