Julian-Chávez Brenda, Siqueiros-Cendón Tania S, Torres-Castillo Jorge Ariel, Sinagawa-García Sugey Ramona, Abraham-Juárez María Jazmín, González-Barriga Carmen Daniela, Rascón-Cruz Quintín, Siañez-Estrada Luis Ignacio, Arévalo-Gallegos Sigifredo, Espinoza-Sánchez Edward Alexander
Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitario S/N Nuevo Campus Universitario, Chihuahua 31125, Chihuahua, Mexico.
Instituto de Ecología Aplicada, Universidad Autónoma de Tamaulipas, Ave. División del Golfo 356, Col. Libertad, Ciudad Victoria 87019, Tamaulipas, Mexico.
Insects. 2024 Dec 17;15(12):1000. doi: 10.3390/insects15121000.
In the search for effective strategies to control the Colorado Potato Beetle, RNA interference technology has emerged as a promising method due to its capacity to suppress genes selectively. Factors such as the target gene and double-stranded RNA (dsRNA) length are critical for optimizing gene silencing efficiency. In this study, we designed and synthesized in vitro dsRNAs of varying lengths targeting the gene, which encodes the AChE1 isoform of acetylcholinesterase in the beetle. All tested dsRNA lengths (222 bp, 543 bp, 670 bp, and 870 bp) promoted transcript reduction. The 670 bp dsRNA was the most effective, reducing transcript levels by approximately 40% by day seven, followed by the 543 bp dsRNA. No significant differences were observed between the 222 bp and 870 bp dsRNAs. Furthermore, all of the dsRNA lengths resulted in reduced weight gain and increased mortality in larvae, with the 670 bp dsRNA showing the highest mortality rate, leaving only 63% larval survival, a trend that persisted through day nine. These findings emphasize that dsRNA length is a key factor in the silencing response, underscoring the importance of selecting the optimal length while considering the gene's target, stability, and delivery methods. This study contributes to establishing design criteria for dsRNA, aiding in the development of more effective and sustainable pest management strategies.
在寻找控制科罗拉多马铃薯甲虫的有效策略过程中，RNA干扰技术因其能够选择性抑制基因的能力而成为一种有前景的方法。诸如靶基因和双链RNA（dsRNA）长度等因素对于优化基因沉默效率至关重要。在本研究中，我们设计并体外合成了针对该基因的不同长度的dsRNA，该基因编码甲虫乙酰胆碱酯酶的AChE1同工型。所有测试的dsRNA长度（222 bp、543 bp、670 bp和870 bp）均促进了转录本减少。670 bp的dsRNA最有效，到第7天转录本水平降低了约40%，其次是543 bp的dsRNA。222 bp和870 bp的dsRNA之间未观察到显著差异。此外，所有dsRNA长度均导致幼虫体重增加减少和死亡率增加，670 bp的dsRNA死亡率最高，幼虫存活率仅为63%，这一趋势持续到第9天。这些发现强调dsRNA长度是沉默反应的关键因素，突出了在考虑基因靶点、稳定性和递送方法时选择最佳长度的重要性。本研究有助于建立dsRNA的设计标准，有助于开发更有效和可持续的害虫管理策略。
