Corteva AgriScience, Johnston, IA, USA.
Methods Mol Biol. 2022;2360:91-103. doi: 10.1007/978-1-0716-1633-8_9.
Identification of active target genes in bioassay screening is the first important step for application of RNA interference (RNAi) for pest control. Here, we describe the methodology for performing high-throughput RNAi target screening against important agriculture pest, Western corn rootworm in 96-well microplate. Two approaches are presented to identify active targets from random-cDNA library or testing a certain group of specific targets via in silico sequence analysis. Methods of PCR primer design, DNA template preparation, and dsRNA production described here can be applied for other pests.
在生物测定筛选中鉴定活性靶基因是应用 RNA 干扰 (RNAi) 进行害虫防治的第一步。在这里,我们描述了针对重要农业害虫——西方玉米根虫在 96 孔微板中进行高通量 RNAi 靶标筛选的方法。提出了两种方法来从随机 cDNA 文库中鉴定活性靶标,或通过计算机序列分析测试特定靶标组。这里描述的 PCR 引物设计、DNA 模板制备和 dsRNA 生产方法可应用于其他害虫。