Method for efficiently and rapidly screening functional sites in gene regulation region and application thereof

The invention discloses a method for efficiently and rapidly screening functional sites in a gene non-coding regulation region and application thereof. The present invention combines Cre/ lox and CRISPR/ Cas9 technologies to introduce in-vitro constructed exogenous mutation into genome so as to reach in-situ mutation effect, and to screen the functional sites of gene non-coding region (such as enhancer) efficiently and quickly. The point mutation is introduced into the regulatory region of genome by CRISPR/ Cas9 technique, although the effect of in-situ mutation can be achieved, the post-screening workload is high. Some mutation sites are not suitable for the design of sgRNA, and are limited by the region of PAM sequence. The method breaks through the limitation and can carry out point mutation at any site in the gene regulation region. In addition, the technology can simultaneously introduce the mutation of two or more sites at the same time, and realize the excision of specific sequences in the gene regulat