期刊
SCIENCE CHINA-LIFE SCIENCES
卷 62, 期 1, 页码 1-7出版社
SCIENCE PRESS
DOI: 10.1007/s11427-018-9402-9
关键词
CRISPR; Cas; genome editing; mutation identification; Hi-TOM
类别
资金
- National Key Research and Development Program of China [2017YFD0102002]
- Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences
- National Natural Science Foundation of China [31401363]
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.
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