Persistent Parental RNAi in the Beetle Tribolium castaneum Involves Maternal Transmission of Long Double-Stranded RNA.
Persistent Parental RNAi in the Beetle Tribolium castaneum Involves Maternal Transmission of Long Double-Stranded RNA.
复制标题
DOI：
10.1002/ggn2.202100064
复制
发表时间：
2022-09
期刊：
影响因子：
--
通讯作者：
Panfilio, Kristen A
中科院分区：
文献类型：
--
作者：
Horn, Thorsten;Narov, Kalin D;Panfilio, Kristen A
亲本RNA干扰（pRNAi）是一种功能强大且广泛使用的基因特异性敲除方法。然而，在昆虫中，它的功效因物种而异，并且系统反应如何从母亲传递给后代仍然难以捉摸。使用甲虫赤拟谷盗（Tribolium castaneum），报道了区分双链RNA（dsRNA）与内源mRNA的存在的RT-qPCR策略。发现注射的dsRNA直接传递到卵中并在整个胚胎发生中持续存在。尽管来自母体的dsRNA有这种消耗，但显示强pRNAi可以持续数月，然后以菌株特异性速率减弱。在寻找细胞摄取长dsRNA进入鸡蛋的受体蛋白时，还提出了候选蛋白的cDNA基因组学分析方法。一种可视化策略的基础上分类层次评估的直系聚类数据，以快速评估基因年龄和拷贝数的变化，细化基于序列的证据，证明。因此，记录了节肢动物中SID-1样通道蛋白的重复损失，包括异翅目（真正的昆虫）中的大规模损失，尽管如此，它们对pRNAi高度敏感。总体而言，阐明了在长期系统性敲除dsRNA传递的分子机制的突出问题的背景下昆虫pRNAi的实际考虑。系统性RNA干扰（RNAi），包括治疗母亲敲低后代的基因，是昆虫研究和害虫管理的有力工具。为了阐明分子机制，研究表明长双链RNA在长达数月的敲除过程中直接传递到卵子中。进化的起源和损失的蛋白质参与细胞的摄取是确定的基因组图谱。
Parental RNA interference (pRNAi) is a powerful and widely used method for gene‐specific knockdown. Yet in insects its efficacy varies between species, and how the systemic response is transmitted from mother to offspring remains elusive. Using the beetle Tribolium castaneum, an RT‐qPCR strategy to distinguish the presence of double‐stranded RNA (dsRNA) from endogenous mRNA is reported. It is found that injected dsRNA is directly transmitted into the egg and persists throughout embryogenesis. Despite this depletion of dsRNA from the mother, it is shown that strong pRNAi can persist for months before waning at strain‐specific rates. In seeking the receptor proteins for cellular uptake of long dsRNA into the egg, a phylogenomics profiling approach of candidate proteins is also presented. A visualization strategy based on taxonomically hierarchical assessment of orthology clustering data to rapidly assess gene age and copy number changes, refined by sequence‐based evidence, is demonstrated. Repeated losses of SID‐1‐like channel proteins in the arthropods, including wholesale loss in the Heteroptera (true bugs), which are nonetheless highly sensitive to pRNAi, are thereby documented. Overall, practical considerations for insect pRNAi against a backdrop of outstanding questions on the molecular mechanism of dsRNA transmission for long‐term, systemic knockdown are elucidated. Systemic RNA interference (RNAi), including treating the mother to knock down genes in offspring, is a powerful tool in insects for research and pest management. To clarify molecular mechanisms, it is shown that long double‐stranded RNA is transmitted directly into eggs during months‐long knockdown. Evolutionary origin and loss of proteins implicated in cellular uptake is ascertained by phylogenomic profiling.