Synthetic CRISPR-Cas9 gene drive has been developed to control harmful species. However, resistance to Cas9 gene drive can be acquired easily when DNA repair mechanisms patch up the genetic insults introduced by Cas9 and incorporate mutations to the sgRNA target. Although many strategies to reduce the occurrence of resistance have been developed so far, they are difficult to implement and not always effective. Here, Cas12f1, a recently developed CRISPR-Cas system with minimal potential for causing mutations within target sequences, has been explored as a potential platform for yielding low-resistance in gene drives. We construct Cas9 and Cas12f1 gene drives in a fast-replicating DNA virus, HSV1. Cas9 and Cas12f1 gene drives are able to spread among the HSV1 population with specificity towards their target sites, and their transmission among HSV1 viruses is not significantly affected by the reduced fitness incurred by the viral carriers. Cas12f1 gene drives spread similarly as Cas9 gene drives at high introduction frequency but transmit more slowly than Cas9 gene drives at low introduction frequency. However, Cas12f1 gene drives outperform Cas9 gene drives because they reach higher penetration and induce lower resistance than Cas9 gene drives in all cases. Due to lower resistance and higher penetration, Cas12f1 gene drives could potentially supplant Cas9 gene drives for population control.

中文翻译：

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Cas12f1 基因驱动在疱疹病毒中有效繁殖，诱导的耐药性最小


合成 CRISPR-Cas9 基因驱动已被开发用于控制有害物种。然而，当 DNA 修复机制修补 Cas9 引入的遗传损伤并将突变整合到 sgRNA 靶标时，很容易获得对 Cas9 基因驱动的耐药性。尽管迄今为止已经开发了许多减少耐药性发生的策略，但它们难以实施且并不总是有效。在这里，Cas12f1 是一种最近开发的 CRISPR-Cas 系统，在靶序列内引起突变的可能性最小，已被探索为在基因驱动中产生低抗性的潜在平台。我们在快速复制的 DNA 病毒 HSV1 中构建 Cas9 和 Cas12f1 基因驱动。Cas9 和 Cas12f1 基因驱动能够在 HSV1 人群中传播，对其目标位点具有特异性，并且它们在 HSV1 病毒中的传播不会受到病毒携带者降低的适应性的显着影响。Cas12f1 基因驱动器的传播方式与高引入频率下的 Cas9 基因驱动相似，但比低引入频率下的 Cas9 基因驱动传播得慢。然而，Cas12f1 基因驱动的性能优于 Cas9 基因驱动，因为它们在所有情况下都比 Cas9 基因驱动达到更高的渗透率和更低的耐药性。由于较低的抗性和较高的渗透性，Cas12f1 基因驱动可能会取代 Cas9 基因驱动进行种群控制。