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产卵后冷却延长了Bombyx mori中微注射介导基因组修改的允许期

Keiro Uchino¹, Ryusei Waizumi¹, Megumi Sumitani¹

¹Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Owashi, Tsukuba 305-8634, Ibaraki, Japan.

International journal of molecular sciences

|December 17, 2024

概括

在微注射之前冷却丝虫蛋显著提高了转基因效率. 这种蛋冷却处理提高了昆虫基因组修改率,对于发展阶段短的物种非常有用.

科学领域:

* 发育生物学发育生物学
* 遗传学遗传学是一门学科
* 昆虫学昆虫学

背景情况:

*昆虫的转基因效率高度依赖于用于微注射的卵的发育阶段.
*优化卵子发育状态对于像丝虫,Bombyx mori等物种的成功基因组修改至关重要.

研究的目的:

* 调查卵冷却对Bombyx mori*转基因效率的影响.
* 为了确定通过冷却改变卵子发育状态是否会增加转基因发生的频率.

主要方法:

*在25°C温度下微注射各种发育阶段的*Bombyx mori*蛋 (产卵后4,8,12小时).
*微注射在不同发育阶段的Bombyx mori*蛋,冷却至10°C5小时或24小时.
*基因淘汰实验以冷却卵子为目标,针对BmBLOS2*基因.

主要成果:

*在产卵后4小时的微注射 (hAEL) 显示效率比8hAEL高5倍;在25°C的12hAEL没有发生转基因.
*将蛋冷却到10°C,使得5hAEL和24hAEL的转基因发生成功.
*冷却后的蛋显示出BmBLOS2的基因淘汰表型,即使在G0注射期间48haEL.

结论:

* 蛋冷处理显著提高了昆虫基因组修改的效率.
* 这种方法提供了一种可行的策略来提高转基因发生率,特别是在短暂的突触性皮质前阶段的昆虫中.

关键词:

波姆比克斯 (Bombyx mori mori mori) 是一个有趣的动物.发育值温度是指发育值温度.微注射微注射的方法微注射的允许期为微注射.

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