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bioRxiv · 10.1101/235903

Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems

Liu, Q.·Wang, C.·Jiao, X.·Zhang, H.·Song, L.·Li, Y.·Gao, C.·Wang, K.
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Abstract

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 x N (N denotes the number of targets) genome-modified sites. The strategy simplified and streamlined the process of next-generation sequencing (NGS) library construction by fixing the bridge sequences and barcoding 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. 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 that frequently induced by genome editing. Hi-TOM does not require specially 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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Liu, Q., Wang, C., Jiao, X., Zhang, H., Song, L., Li, Y., Gao, C., Wang, K.. 2017-12-18. Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems. https://doi.org/10.1101/235903

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