Córdoba Lourdes E, Pérez de Rosas Alicia R, García Beatriz A, Stroppa María M
Instituto de Investigaciones en Ciencias de la Salud (INICSA), CONICET and Cátedra de Bioquímica y Biología Molecular, Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
Instituto de Investigaciones en Ciencias de la Salud (INICSA), CONICET and Cátedra de Bioquímica y Biología Molecular, Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
STAR Protoc. 2025 Jun 20;6(2):103865. doi: 10.1016/j.xpro.2025.103865. Epub 2025 May 30.
RNA interference (RNAi) is a technique that enables post-transcriptional gene silencing in organisms. Here, we present a protocol for silencing the clock gene period (per) in adult insects of Triatoma infestans using RNAi. We describe steps for insect rearing, synthesis of double-stranded RNA (dsRNA) of target genes, and abdominal microinjection with dsRNA. We then detail procedures for collecting and extracting RNA from heads at different time points and evaluating silencing efficiency using quantitative reverse-transcription PCR (RT-qPCR). For complete details on the use and execution of this protocol, please refer to Córdoba et al..
RNA干扰(RNAi)是一种能够在生物体中实现转录后基因沉默的技术。在此,我们展示了一种使用RNAi使成年克氏锥蝽中生物钟基因周期(per)沉默的方案。我们描述了昆虫饲养、靶基因双链RNA(dsRNA)合成以及dsRNA腹部显微注射的步骤。然后,我们详细说明了在不同时间点从头部收集和提取RNA以及使用定量逆转录PCR(RT-qPCR)评估沉默效率的程序。有关此方案使用和执行的完整详细信息,请参考科尔多瓦等人的研究。