RNA Interference Method for Gene Function Analysis in the Japanese Rhinoceros Beetle Trypoxylus dichotomus

Kazuki Sakura; Shinichi Morita; Teruyuki Niimi

doi:10.21769/BioProtoc.4396

RNA Interference Method for Gene Function Analysis in the Japanese Rhinoceros Beetle Trypoxylus dichotomus

Bio Protoc. 2022 Apr 20;12(8):e4396. doi: 10.21769/BioProtoc.4396.

Authors

Kazuki Sakura¹, Shinichi Morita¹, Teruyuki Niimi^{1

2}

Affiliations

¹ Division of Evolutionary Developmental Biology, National Institute for Basic Biology, 38 Nishigonaka, Myodaiji, Okazaki, Japan.
² Department of Basic Biology, School of Life Science, The Graduate University for Advanced Studies, SOKENDAI, 38 Nishigonaka, Myodaiji, Okazaki, Japan.

Abstract

In the Japanese rhinoceros beetle Trypoxylus dichotomus, various candidate genes required for a specific phenotype of interest are listed by next-generation sequencing analysis. Their functions were investigated using RNA interference (RNAi) method, the only gene function analysis tool for T. dichotomusdeveloped so far. The summarized procedure for the RNAi method used for T. dichotomusis to synthesize double-stranded RNA (dsRNA), and inject it in larvae or pupae of T. dichotomus. Although some dedicated materials or equipment are generally required to inject dsRNA in insects, the advantage of the protocol described here is that it is possible to inject dsRNA in T. dichotomuswith one syringe.

Keywords: Double-stranded RNA; Gene knock-down; Japanese rhinoceros beetle; RNA interference; Trypoxylus dichotomus.