CRISPR-based synthetic gene drives have the potential to deliver a more effective and humane method of invasive vertebrate pest control than current strategies. Relatively efficient CRISPR gene drive systems have been developed in insects and yeast but not in mammals. Here, we investigated the efficiency of CRISPR-Cas9-based gene drives in Mus musculus by constructing “split drive” systems where gRNA expression occurs on a separate chromosome to Cas9, which is under the control of either a zygotic (CAG) or germline (Vasa) promoter. While both systems generated double-strand breaks at their intended target site in vivo, no homology-directed repair between chromosomes (“homing”) was detectable. Our data indicate that robust and specific Cas9 expression during meiosis is a critical requirement for the generation of efficient CRISPR-based synthetic gene drives in rodents.

中文翻译：

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小鼠合子和种系基因驱动的进展

基于 CRISPR 的合成基因驱动有可能提供一种比当前策略更有效、更人道的侵入性脊椎动物害虫控制方法。相对有效的 CRISPR 基因驱动系统已在昆虫和酵母中开发，但尚未在哺乳动物中开发。在这里，我们研究了基于CRISPR-Cas9基因驱动器的效率小家鼠通过其中gRNA表达一个单独的染色体Cas9，其或者是一个合子（CAG）或种系的控制下（上发生构建“分裂驱动”系统瓦萨）发起人。虽然这两个系统都在体内预期的目标位点产生了双链断裂，没有检测到染色体之间的同源定向修复（“归巢”）。我们的数据表明，减数分裂过程中强大且特异性的 Cas9 表达是在啮齿动物中产生有效的基于 CRISPR 的合成基因驱动的关键要求。