Generating single-sex litters: development of CRISPR-Cas9 genetic tools to produce all-male offspring
Generating single-sex litters: development of CRISPR-Cas9 genetic tools to produce all-male offspring
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DOI：
10.1101/2020.09.07.285536
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发表时间：
2020-09
期刊：
影响因子：
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通讯作者：
C. Douglas;Valdone Maciulyte;Jasmin Zohren;Daniel M. Snell;O. Ojarikre;P. Ellis;James M. A. Turner
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文献类型：
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作者：
C. Douglas;Valdone Maciulyte;Jasmin Zohren;Daniel M. Snell;O. Ojarikre;P. Ellis;James M. A. Turner
动物在科学和全球农业资源中是非常有用的遗传工具。然而，一个单一的性别往往是多余的，目前的遗传方法生产全女性或全男性窝是低效的。使用小鼠作为模型，我们开发了一种合成的，两部分的双组分策略，用于产生全雄性窝仔。我们使用CRISPR-Cas9基因组编辑技术在常染色体和X染色体上产生大的稳定敲入。双组分系统通过靶向必需拓扑异构酶1基因的Cas9转基因和sgRNA转基因的性别特异性共遗传发挥作用。这项技术被证明在产生靶向突变方面非常有效，导致目标性别的胚胎死亡。我们的研究是第一个使用CRISPR-Cas9双组分系统成功产生全雄性哺乳动物窝仔的研究，并为实验室或农业研究产生单性别窝仔提供了巨大的进步。
Animals are extremely useful genetic tools in science and global resources in agriculture. However, a single sex is often required in surplus, and current genetic methods for producing all-female or all-male litters are inefficient. Using the mouse as a model, we developed a synthetic, two-part bicomponent strategy for generating all-male litters. We achieved this using CRISPR-Cas9 genome editing technology to generate large stable knock-ins on the autosomes and X chromosome. The bicomponent system functions via the sex-specific co-inheritance of a Cas9 transgene and an sgRNA transgene targeting the essential Topoisomerase 1 gene. This technology proved to be highly efficient in generating on-target mutations, resulting in embryonic lethality of the target sex. Our study is the first to successfully generate all-male mammalian litters using a CRISPR-Cas9 bicomponent system and provides great strides towards generating single-sex litters for laboratory or agricultural research.