The free-living nematode C. elegans remains one of the most robust and flexible genetic systems for interrogating the complexities of animal biology. Targeted genetic manipulations, such as RNA interference (RNAi), CRISPR or array-based transgenesis, all rely on initial delivery of nucleic acids. Delivery of dsRNA by feeding can be effective, but expression in E. coli is not conducive to experiments intended to remain sterile or with defined microbial communities. Soaking-based delivery requires prolonged exposure of animals to high material concentrations without a food source and is of limited throughput. Last, microinjection of individual animals can precisely deliver materials to animals' germlines, but is limited by the need to target and inject each animal one-by-one. Thus, we sought to address some of these challenges in nucleic acid delivery by developing a population-scale delivery method. We demonstrate efficient electroporation-mediated delivery of dsRNA throughout the worm and effective RNAi-based silencing, including in the germline. Finally, we show that guide RNA delivered by electroporation can be utilized by transgenic Cas9 expressing worms for population-scale genetic targeting. Together, these methods expand the scale and scope of genetic methodologies that can be applied to the C. elegans system.

中文翻译：

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通过电穿孔将种群规模的核酸递送至秀丽隐杆线虫

自由生活的线虫秀丽隐杆线虫仍然是最健壮和灵活的遗传系统之一，用于询问动物生物学的复杂性。靶向遗传操作，例如RNA干扰（RNAi），CRISPR或基于阵列的转基因，都依赖于核酸的初始递送。通过喂养递送dsRNA可能是有效的，但在大肠杆菌中表达不利于旨在保持无菌或具有特定微生物群落的实验。基于浸泡的递送需要动物在没有食物源的情况下长时间暴露于高物质浓度，并且产量有限。最后，对单个动物进行显微注射可以将材料精确地传递到动物的种系中，但是由于需要逐一靶向和注射每个动物而受到限制。因此，我们试图通过开发群体规模的递送方法来解决核酸递送中的一些挑战。我们展示了整个蠕虫中有效的电穿孔介导的dsRNA传递以及包括种系在内的基于RNAi的有效沉默。最后，我们表明通过电穿孔传递的指导RNA可以被转基因Cas9表达蠕虫用于人群规模的遗传靶向。一起，秀丽隐杆线虫系统。