CRISPR‐based gene drives bias inheritance in their favour by inducing double‐stranded breaks (DSBs) at wild‐type homologous loci and using the drive transgene as a repair template—converting drive heterozygotes into homozygotes. Recent studies have shown that alternate end‐joining repair mechanisms produce cut‐resistant alleles that rapidly induce drive failure. Multiplexing—simultaneously targeting multiple sites at the wild‐type locus—is commonly assumed to overcome this issue since resistance would need to develop at all target sites for the system to fail. This may work for some population suppression drives targeting essential (e.g. viability or fertility) genes if careful design can ensure cut‐resistant alleles themselves have low fitness. However, here, models are used to demonstrate that this approach will be ineffective when targeting neutral loci. We then go on to compare the performance of four alternative population‐level multiplexing approaches with standard individual‐level multiplexing. Two of these approaches have mechanisms preventing them from becoming linked, thus avoiding multiple simultaneous DSBs and giving a large improvement. Releasing multiple unlinked drives gives a modest improvement, while releasing multiple drives that may become linked over time produces a decrease in performance under the conditions tested here. Based on performance and technical feasibility, we then take one approach forward for further investigation, demonstrating its robustness to different performance parameters and its potential for controlling very large target populations.

中文翻译：

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群体水平多重化：管理针对中性位点的基因驱动抗性进化的有前途的策略。

基于 CRISPR 的基因通过在野生型同源位点诱导双链断裂 (DSB) 并使用驱动转基因作为修复模板，将驱动杂合子转化为纯合子，从而驱动有利于它们的偏向遗传。最近的研究表明，替代末端连接修复机制会产生抗切割等位基因，从而迅速导致驱动器故障。通常认为多重化（同时针对野生型基因座的多个位点）可以克服这个问题，因为需要在所有目标位点上产生抗性，系统才会失败。如果仔细的设计可以确保抗切割等位基因本身具有较低的适应性，这可能适用于某些针对必需（例如生存力或生育力）基因的群体抑制驱动力。然而，这里使用模型来证明这种方法在针对中性基因座时无效。然后，我们继续将四种替代群体水平多路复用方法与标准个体水平多路复用方法的性能进行比较。其中两种方法具有防止它们相互链接的机制，从而避免多个同时 DSB 并给出了很大的改进。释放多个未链接的驱动器会带来一定的改进，而释放可能随着时间的推移而链接的多个驱动器会在此处测试的条件下导致性能下降。基于性能和技术可行性，我们随后采取一种方法进行进一步研究，证明其对不同性能参数的鲁棒性及其控制非常大的目标群体的潜力。