Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems
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 describe a cheap, practicable and high-throughput screening strategy that allows parallel screening of 96 × N (N denotes the number of targets) genome-modified sites. The strategy simplified the construction of next-generation sequencing (NGS) library by fixing the second-round PCR primers. We also developed Hi-TOM (available at http://www.hi-tom.net/hi-tom/), an online tool to track the mutations with precise percentage. Hi-TOM does not require cumbersome parameter configuration or additional data analysis; thus, it can be exploited by researchers who are unfamiliar with NGS or bioinformatics. Analysis of the samples from rice, hexaploid wheat and human cells reveals that the Hi-TOM tool has high reliability and sensitivity. The simplicity, convenience and comprehensive output make Hi-TOM particularly suitable for high-throughput identification of all types of mutations induced by CRISPR/Cas systems.
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