High-throughput identification method for genome insulator

Abstract

The invention discloses a high-throughput identification method for a genome insulator. The high-throughput identification method for the genome insulator includes the steps of (1) building a reportervector, specifically, the reporter vector sequentially comprises a promoter, an ORF region, an intron, an insulator candidate sequence insertion region and an enhancer, the enhancer is used for enhancing expression of an upstream sequence, and an insulator candidate sequence is inserted in the insulator candidate sequence insertion region; (2) expressing and transcribing the reporter vector; and(3) detecting the transcribing condition of the edited transcribing sequence, and determining whether the insulator candidate sequence is the insulator or not. A reporter vector library built by the method can cover 90-99% of genome noncoding regions, protein position information is not needed, and overall analysis is achieved; the built library is transferred into a cell, whether nearly the wholegenomic sequence has insulator activity or not can be detected at the same time, large-scale parallel analysis can be carried out, and single and small-quantity analysis is avoided; and since the built library covers nearly all sites, zones of ignorance can be also detected and analyzed.

Description

technical field [0001] The invention relates to genomics, in particular to a high-throughput identification method for genome isolators. Background technique [0002] With the completion of the Human Genome Project and the formation and maturity of the concept of precision medicine, people have more and more comprehensive requirements for the knowledge of the function of the genome itself. However, in fact, scientists' understanding of the function of the genome lags far behind the actual needs. , so that the current precision medicine is more of a concept and it is difficult to implement it, or the foundation for implementation is unstable. [0003] In addition to genes, there is a serious lack of functional knowledge about non-coding sequences that account for 98% of the human genome. How to analyze and determine the potential functions of genome sequences is a major theoretical issue in biology and the basis for applications. Isolator regulatory elements are an important...

AI Technical Summary

Problems solved by technology

[0005] First, rely on the existing insulator protein binding site information to predict, if there is no protein binding information, it cannot be predicted;

[0006] Second, whether the predicted site has a function can only be determined individually and cannot be analyzed on a large scale;

[0007] Third, the presence or absence of insulator function in regions bound by non-isolator proteins can neither be predicted nor analysed.

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