Method for detecting gene editing efficiency and gene editing mode and application of method

Abstract

The invention relates to a method for detecting gene editing efficiency and a gene editing mode and an application of the method, and belongs to the technical field of gene detection. The method comprises the following steps: S1, extracting genomic DNA of a sample, designing primers for a to-be-detected region, and amplifying the primers; S2, constructing a library to perform high-throughput sequencing, and filtering, splitting and splicing the data to obtain a sequence of the to-be-detected region; S3, comparing the sequence of the to-be-detected region with a genome reference sequence to obtain the gene editing efficiency and mode of each sample; and S4, performing significance analysis on the gene editing efficiency of processing group samples and control group samples, and outputting acorrected result. Compared with the prior art, the method has the advantages that the gene library construction process is simplified, the data analysis and processing speed is increased, the economy, sensitivity, specificity and accuracy of gene editing efficiency detection are improved, and multi-dimensional evaluation of a gene editing mode is also provided.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method for detecting gene editing efficiency and gene editing mode and its application. Background technique [0002] Gene editing technology has been widely used, showing great potential in gene research, gene therapy and genetic improvement. After gene editing, target DNA analysis of gene editing mutants is required to understand gene editing efficiency and pattern. In the field of gene detection technology, the T7 endonuclease digestion method and the TA clone Sanger sequencing method were used early to estimate the gene editing efficiency. The detection throughput of these two methods is very low, and the detection process is time-consuming, laborious and expensive. The identification results are not very accurate. Currently more advanced genetic testing methods are based on high-throughput sequencing. This method has high throughput, and its sensitivity and speci...

AI Technical Summary

Problems solved by technology

[0003] However, the gene editing analysis platform based on high-throughput sequencing has always had several major defects: 1) The preparation process of high-throughput sequencing library is complicated, which increases the time cost and economic cost of platform operation; 2) The large amount of high-throughput sequencing data, The process of data processing and analysis is complex, requiring high equipment and taking a long time; 3) Due to the high sensitivity of the high-throughput sequencing platform, its false positive rate is also high, and the traditional analysis and processing process cannot well exclude false positive samples; 4) Traditional analysis methods can only output gene editing efficiency, but cannot characterize gene editing patterns well

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