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© The Author(s) 2025. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.

Abstract

The Japanese rhinoceros beetle, Trypoxylus dichotomus, possesses large horns on its head and thorax, features whose biological significance has been explored across various fields, including evolutionary developmental biology, behavioral ecology, and materials science. To investigate the molecular basis of these characteristics, systemic larval RNA interference (RNAi) has been employed as a primary loss-of-function genetic tool. However, gain-of-function analyses and region-specific gene function assessments remain underdeveloped, thereby limiting the comprehensive understanding of the molecular mechanisms involved. To address this limitation, we developed an in vivo electroporation technique to introduce exogenous DNA vectors directly into the somatic tissues of T. dichotomus larvae to express the genes of interest. Additionally, we utilized the piggyBac transposon system to insert the exogenous DNA vectors into the host genome for stable gene expression. Our findings indicate that the T. dichotomus actin A3 gene promoter exhibits sufficient transcriptional activity in the early postembryonic stage of T. dichotomus via electroporation. Furthermore, we observed that this promoter functions effectively across a diverse range of insect species, including the harlequin ladybug, Harmonia axyridis and the silkworm, Bombyx mori, suggesting the broad applicability of the T. dichotomus actin A3 promoter in various insects.

Details

Title
Electroporation-mediated functional analysis method of genes in the giant insect Trypoxylus dichotomus
Author
Sakura, Kazuki 1 ; Morita, Shinichi 2 ; Ando, Toshiya 3 ; Yuzaki, Karen 4 ; Shigenobu, Shuji 5 ; Niimi, Teruyuki 6 

 National Institute for Basic Biology, Division of Evolutionary Developmental Biology, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593) 
 National Institute for Basic Biology, Division of Evolutionary Developmental Biology, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593); The Graduate University for Advanced Studies, SOKENDAI, Basic Biology Program, Okazaki, Japan (GRID:grid.275033.0) (ISNI:0000 0004 1763 208X) 
 National Institute for Basic Biology, Division of Evolutionary Developmental Biology, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593); Kyoto University, The Hakubi Center for Advanced Research, Kyoto, Japan (GRID:grid.258799.8) (ISNI:0000 0004 0372 2033) 
 National Institute for Basic Biology, Division of Evolutionary Developmental Biology, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593); Nagoya University, Graduate School of Bioagricultural Sciences, Chikusa, Nagoya, Japan (GRID:grid.27476.30) (ISNI:0000 0001 0943 978X) 
 The Graduate University for Advanced Studies, SOKENDAI, Basic Biology Program, Okazaki, Japan (GRID:grid.275033.0) (ISNI:0000 0004 1763 208X); National Institute for Basic Biology, Laboratory of Evolutionary Genomics, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593); National Institute for Basic Biology, Trans-Omics Facility, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593) 
 National Institute for Basic Biology, Division of Evolutionary Developmental Biology, Okazaki, Japan (GRID:grid.419396.0) (ISNI:0000 0004 0618 8593); The Graduate University for Advanced Studies, SOKENDAI, Basic Biology Program, Okazaki, Japan (GRID:grid.275033.0) (ISNI:0000 0004 1763 208X); Nagoya University, Graduate School of Bioagricultural Sciences, Chikusa, Nagoya, Japan (GRID:grid.27476.30) (ISNI:0000 0001 0943 978X) 
Pages
25923
Publication year
2025
Publication date
2025
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
3231090406
Copyright
© The Author(s) 2025. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.