BACKGROUNDSex control technology has great potential to improve the efficiency of economic insect utilization and integrated pest management by enabling targeted population suppression or elimination. The silkworm (Bombyx mori), a lepidopteran model, holds immense economic importance attributable to its unique silk production capabilities. Male silkworms favored over females as a result of their higher yield and quality silk. Doublesex (dsx) is a key gene for sex determination in insects. Previously, inactivation of Bmdsx, among other sex determination‐related genes, resulted in intersex individuals rather than a complete female‐to‐male transformation.RESULTSWe used CRISPR/Cas9 technology to edit the splice sites of Bmdsx at female‐specific exons 3 and 4 to induce transformation of BmdsxF to BmdsxM. The edited genome produced only male‐specific BmdsxM. The results revealed that these genetic modifications had significant effects on genital development in female individuals. The edited genome led to the emergence of male‐specific gonads in females, causing genital developmental defects. However, it is worth noting that males remained unaffected by these genetic alterations. Furthermore, we conducted a comprehensive analysis of dsx gene structure and explored the potential applicability of this method for inducing female‐to‐male transgenic transformation in other insects.CONCLUSIONSThis study demonstrates that precision editing of dsx splice sites can enforce unidirectional sex conversion in insects, disrupting female reproductive capacity without affecting males. Overall, these findings provide valuable insights into our understanding of gonadal development and insect population management. © 2025 Society of Chemical Industry.

中文翻译：

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家蚕中的双性基因：CRISPR 介导的通过外显子跳跃的雌性到雄性逆转

背景 防治技术具有巨大的潜力，可以通过实现有针对性的种群抑制或消除来提高经济昆虫利用和综合害虫管理的效率。蚕 （Bombyx mori） 是一种鳞翅目模型，由于其独特的丝绸生产能力而具有巨大的经济意义。雄蚕比雌蚕更受青睐，因为它们的产量和质量更高。Doublesex （dsx） 是昆虫性别决定的关键基因。以前，Bmdsx 以及其他性别决定相关基因的失活导致双性人个体，而不是完全的女性到男性转化。结果我们使用 CRISPR/Cas9 技术编辑 Bmdsx 在女性特异性外显子 3 和 4 处的剪接位点，以诱导 BmdsxF 向 BmdsxM 的转化。编辑后的基因组仅产生雄性特异性 BmdsxM。结果显示，这些基因改造对女性个体的生殖器发育有显着影响。编辑后的基因组导致女性出现男性特异性性腺，导致生殖器发育缺陷。然而，值得注意的是，雄性仍然不受这些基因改变的影响。此外，我们对 dsx 基因结构进行了全面分析，并探讨了该方法在其他昆虫中诱导雌性转雄性转基因的潜在适用性。结论该研究表明，对 dsx 剪接位点的精确编辑可以强制昆虫进行单向性别转换，破坏雌性生殖能力而不影响雄性。总的来说，这些发现为我们对性腺发育和昆虫种群管理的理解提供了有价值的见解。© 2025 化工学会.