Whole genome off-target evaluation method and application thereof

Abstract

The invention provides a method for evaluating genome off-target, which comprises the following steps: carrying out methylation and histone modification on a targeting site by using an epigenetic editing tool, and carrying out whole genome transcriptome sequencing, whole genome methylation sequencing and CUTamp on the modified gene, carrying out Tag sequencing, wherein CUTamp is used as a template; tag sequencing comprises H3K4me3, H3K9ac, H3K27ac, H3K9me3 and H3K27me3 histone modification detection, and a whole transcriptome quantitative result, a whole genome methylation level quantitative result and CUTamp are obtained; tag peak abundance quantitative results, determining a target site change map, and determining methylation and histone change directions; and finally, carrying out difference analysis on the sequencing result to determine off-target information.

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