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Background <p><i>Aedes albopictus</i> is a primary vector for the transmission of dengue fever. RNA interference (RNAi)-based biocidal technology represents an important alternative and complement to conventional chemically synthesized insecticides.</p> Methods <p><i>Chlorella vulgaris</i> was used as a carrier organism for RNAi-mediated gene silencing of <i>Ae. albopictus</i>. Short hairpin RNA (shRNA) expression vectors targeting the <i>vgat</i> and <i>vmat</i> genes of <i>Ae. albopictus</i> were constructed and subsequently used to transform <i>C. vulgaris</i>. The shRNA-transformed <i>Chlorella</i> were then administered to <i>Ae. albopictus</i> larvae or incorporated into attractive toxic sugar baits (ATSB) for adult feeding. The effects of silencing the <i>vgat</i> and <i>vmat</i> genes on <i>Ae. albopictus</i> were then investigated.</p> Results <p>Both <i>vgat</i> and <i>vmat</i> shRNA-recombinant <i>Chlorella</i> exhibited high lethality against <i>Ae. albopictus</i> larvae and adults. Conversely, shRNA-recombinant <i>Chlorella</i> ATSBs were found to have no lethal effects on non-target organisms, including <i>Drosophila melanogaster</i>, <i>Megalurothrips usitatus</i>, <i>Messor structor</i>, and <i>Lithobates catesbeiana</i> tadpoles. Together, these results confirm the specificity and safety of using shRNA-recombinant <i>Chlorella</i> as a mosquito-killing agent. The results of a semi-field trial demonstrate that recombinant <i>Chlorella</i> ATSB maintained high lethality against <i>Ae. albopictus</i> and significantly reduced the number of eggs laid by this species. Additional experiments revealed that knockdown of the <i>vgat</i> gene in <i>Ae. albopictus</i> resulted in reduced sleep duration, leg twitching, and impaired flight. Conversely, knockdown of the <i>vmat</i> gene increased sleep duration, impaired walking and flight, and induced insensitivity to light levels.</p> Conclusions <p>Recombinant <i>Chlorella</i> expressing <i>vgat</i> and <i>vmat</i> shRNAs demonstrated strong lethality, high specificity, and good safety against both larval and adult <i>Ae. albopictus</i>. Oral delivery of shRNAs effectively knocked down mosquito target genes, providing a convenient approach for functional studies of mosquito genetics. Notably, this study is an important addition to the use of recombinant microalgae as a mosquito biocide.</p> Graphical Abstract <p></p>

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Control of Aedes albopictus populations by silencing of the vesicular GABA transporter (vgat) and the vesicular monoamine transporter (vmat) genes using recombinant Chlorella shRNA

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月14日

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  • Xiaodong Deng,
  • Chunmei Xue,
  • Jingwen Gao,
  • Xiaodan Huang,
  • Dianlong Xu,
  • Juncai Li,
  • Xiaowen Fei

摘要

Background

Aedes albopictus is a primary vector for the transmission of dengue fever. RNA interference (RNAi)-based biocidal technology represents an important alternative and complement to conventional chemically synthesized insecticides.

Methods

Chlorella vulgaris was used as a carrier organism for RNAi-mediated gene silencing of Ae. albopictus. Short hairpin RNA (shRNA) expression vectors targeting the vgat and vmat genes of Ae. albopictus were constructed and subsequently used to transform C. vulgaris. The shRNA-transformed Chlorella were then administered to Ae. albopictus larvae or incorporated into attractive toxic sugar baits (ATSB) for adult feeding. The effects of silencing the vgat and vmat genes on Ae. albopictus were then investigated.

Results

Both vgat and vmat shRNA-recombinant Chlorella exhibited high lethality against Ae. albopictus larvae and adults. Conversely, shRNA-recombinant Chlorella ATSBs were found to have no lethal effects on non-target organisms, including Drosophila melanogaster, Megalurothrips usitatus, Messor structor, and Lithobates catesbeiana tadpoles. Together, these results confirm the specificity and safety of using shRNA-recombinant Chlorella as a mosquito-killing agent. The results of a semi-field trial demonstrate that recombinant Chlorella ATSB maintained high lethality against Ae. albopictus and significantly reduced the number of eggs laid by this species. Additional experiments revealed that knockdown of the vgat gene in Ae. albopictus resulted in reduced sleep duration, leg twitching, and impaired flight. Conversely, knockdown of the vmat gene increased sleep duration, impaired walking and flight, and induced insensitivity to light levels.

Conclusions

Recombinant Chlorella expressing vgat and vmat shRNAs demonstrated strong lethality, high specificity, and good safety against both larval and adult Ae. albopictus. Oral delivery of shRNAs effectively knocked down mosquito target genes, providing a convenient approach for functional studies of mosquito genetics. Notably, this study is an important addition to the use of recombinant microalgae as a mosquito biocide.

Graphical Abstract