ISDT:a Tool for Detecting Exogenous Sequence Insertion Sites based on High-throughput Sequencing Data
Accurate identification of exogenous sequence insertion sites is crucial for transgenic research,gene editing studies,and ex-ploration of disease mechanisms.With the rapid development and widespread application of high-throughput sequencing technology,an increasing number of researchers are applying this technology to the analysis of insertion sites,delving into exogenous sequence inser-tion events.Based on this background,we developed a detection tool for precise identification of exogenous sequence insertion sites based on high-throughput sequencing data(insertion sites detection tool,ISDT).This tool introduces a novel algorithm for precise identification of insertion sites,which relies on alignment of sliding candidate reads and four mode searches(Forward-Reverse mode,Reverse-Forward mode,Forward-Forward mode,and Reverse-Reverse mode)to achieve accurate insertion site identification.To evalu-ate the analytical performance of this insertion site analysis tool,we not only used simulated high-throughput sequencing data containing different types of insertions(including single-point insertion,single exogenous fragment insertion,exogenous homologous fragment in-sertion,exogenous fragment insertion on different chromosomes,exogenous homologous fragment insertion on different chromosomes)and real insertion site high-throughput sequencing data of plant T-DNA and animal F8 gene for comprehensive testing,but also com-pared this analysis tool with published analysis methods.Our tool demonstrated excellent accuracy and sensitivity.The insertion site detection tool developed in this paper,as an efficient and accurate tool for precise identification of insertion sites,offers cross-species applicability and the ability to handle real complex data.This provides significant support for future transgenic research,gene editing studies,and the exploration of disease mechanisms,holding broad application prospects.
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