Objective

With the widespread application of CRISPR/Cas9 gene editing technology, new methods are needed to screen mutants quickly and effectively. Here, we aimed to develop a simple and cost-effective method to screen CRISPR/Cas9-induced mutants.

Result

We report a novel method to identify CRISPR/Cas9-induced mutants through a DNA-guided Argonaute nuclease derived from the archaeon Pyrococcus furiosus. We demonstrated that the Pyrococcus furiosus Argonaute (PfAgo)-based method could distinguish among biallelic mutants, monoallelic mutants and wild type (WT). Furthermore, this method was able to identify 1 bp indel mutations.

Conclusion

The PfAgo-based method is simple to implement and can be applied to screen biallelic mutants and mosaic mutants generated by CRISPR-Cas9 or other kinds of gene editing tools.

中文翻译：

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通过 DNA 引导的 Argonaute 核酸酶快速且经济高效地筛选 CRISPR/Cas9 诱导的突变体

客观的

随着CRISPR/Cas9基因编辑技术的广泛应用，需要新的方法来快速有效地筛选突变体。在这里，我们旨在开发一种简单且具有成本效益的方法来筛选 CRISPR/Cas9 诱导的突变体。

结果

我们报告了一种新方法，通过源自古细菌Pyrococcus furiosus的 DNA 引导的 Argonaute 核酸酶来识别 CRISPR/Cas9 诱导的突变体。我们证明了基于Pyrococcus furiosus Argonaute (PfAgo) 的方法可以区分双等位基因突变体、单等位基因突变体和野生型 (WT)。此外，该方法能够识别 1 bp 插入缺失突变。

结论

基于 PfAgo 的方法实施简单，可用于筛选由 CRISPR-Cas9 或其他类型的基因编辑工具生成的双等位基因突变体和镶嵌突变体。