Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems
Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems
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DOI：
10.1007/s11427-018-9402-9
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发表时间：
2019-01-01
影响因子：
9.1
通讯作者：
Wang, Kejian
中科院分区：
文献类型：
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作者：
Liu, Qing;Wang, Chun;Wang, Kejian
CRISPR/Cas系统已被广泛应用于各种生物体中进行精确的遗传修饰。尽管其重要性和广泛的使用，大规模的突变筛查仍然是耗时，劳动密集型和昂贵的。在这里，我们开发了Hi-TOM（可在https：//doi.org/www.hi-tom.net/hi-tom/获得），这是一种在线工具，用于跟踪多个样品和多个靶位点的精确百分比的突变。我们还描述了相应的下一代测序（NGS）文库构建策略，通过固定桥序列和条形码引物。对水稻、六倍体小麦和人类细胞样本的分析表明，Hi-TOM工具在跟踪各种突变方面具有很高的可靠性和灵敏度，特别是基因组编辑经常诱导的复杂嵌合突变。Hi-TOM不需要条形码引物的特殊设计、繁琐的参数配置或额外的数据分析。因此，精简的NGS文库构建和全面的结果输出使Hi-TOM特别适合于高通量鉴定由CRISPR/Cas系统诱导的所有类型的突变。
The CRISPR/Cas system has been extensively applied to make precise genetic modifications in various organisms. Despite its importance and widespread use, large-scale mutation screening remains time-consuming, labour-intensive and costly. Here, we developed Hi-TOM (available at https://doi.org/www.hi-tom.net/hi-tom/), an online tool to track the mutations with precise percentage for multiple samples and multiple target sites. We also described a corresponding next-generation sequencing (NGS) library construction strategy by fixing the bridge sequences and barcoding primers. Analysis of the samples from rice, hexaploid wheat and human cells reveals that the Hi-TOM tool has high reliability and sensitivity in tracking various mutations, especially complex chimeric mutations frequently induced by genome editing. Hi-TOM does not require special design of barcode primers, cumbersome parameter configuration or additional data analysis. Thus, the streamlined NGS library construction and comprehensive result output make Hi-TOM particularly suitable for high-throughput identification of all types of mutations induced by CRISPR/Cas systems.