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CRISPR/Cas9 Mediated Disruption of Seminal Fluid Protein Sfp62 Induces Male Sterility in Bombyx mori
1Institute of Sericulture and Tea, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Abstract:
Seminal fluid proteins provide factors necessary for development, storage, and activation of sperm. Altered expression of seminal fluid proteins can lead to defect in male infertility. We investigated the function of seminal fluid protein Sfp62 in the model lepidopteran insect Bombyx mori using CRISPR/Cas9 mediated mutagenesis. The knockout of BmSfp62 gene led to male sterility but has no effect on female fertility. The mutation did not affect growth and development of the silkworm of both sexes. Motility of sperm in male mutants was decreased and the mRNA expression levels of other genes encoding seminal fluid proteins were altered in BmSfp62 mutants compared to the wild-type controls. The male sterility caused by mutation of BmSfp62 was stably inherited. As the proteins encoded by Sfp62 genes are conserved among lepidopteran species, Sfp62 is a potential target for the biological management of lepidopteran pests.
Insights
Seminal fluid protein Sfp62 is crucial for male fertility in silkworms. Its absence causes male sterility, impacting sperm motility and offering a potential target for pest control.
Area of Science:
- Entomology
- Genetics
- Reproductive Biology
Background:
- Seminal fluid proteins (SFPs) are vital for sperm function, and their altered expression can cause male infertility.
- Understanding the specific roles of SFPs is essential for addressing reproductive issues and developing novel control strategies.
Purpose of the Study:
- To investigate the function of seminal fluid protein 62 (Sfp62) in the model lepidopteran insect, Bombyx mori.
- To assess the impact of BmSfp62 gene knockout on male fertility, sperm motility, and related gene expression.
Main Methods:
- CRISPR/Cas9 mediated mutagenesis was employed to generate BmSfp62 knockout mutants in Bombyx mori.
- Phenotypic analysis included assessment of male and female fertility, growth, development, and sperm motility.
- mRNA expression levels of other seminal fluid protein genes were analyzed in BmSfp62 mutants.
Main Results:
- BmSfp62 knockout resulted in complete male sterility without affecting female fertility or general growth and development.
- Sperm motility was significantly decreased in male mutants compared to wild-type controls.
- Altered mRNA expression of other seminal fluid protein genes was observed in BmSfp62 mutants.
- The induced male sterility was stably inherited across generations.
Conclusions:
- The seminal fluid protein Sfp62 plays a critical role in male fertility in Bombyx mori.
- BmSfp62 is essential for maintaining normal sperm motility.
- Given the conservation of Sfp62 genes in lepidopteran species, Sfp62 presents a promising target for the biological management of agricultural pests.
