sgRNA for identifying porcine ROSA 26 gene and coding DNA, gene editing method, kit and application thereof

The invention provides sgRNA for identifying a porcine ROSA 26 gene, and coding DNA, a gene editing method, a kit and application thereof, and relates to the technical field of genetic engineering. The sgRNA which specifically recognizes the porcine ROSA 26 gene recognizes a first intron of the porcine ROSA 26 gene, is used for a CRISPR/Cpf 1 system, and comprises a sequence which is responsible for recognizing a target fragment and is shown as SEQ ID NO. 1. The sgRNA is matched with the CRISPR/Cpf 1 system, the knockout efficiency is 27% and the specificity is high; when the sequence knockoutis carried out, the off-target phenomenon can be effectively reduced, and the adverse effect of non-specific cutting on a genome can be reduced. 本发明提供了一种识别猪ROSA26基因的sgRNA及其编码DNA、基因编辑方法、试剂盒和应用，涉及基因工程技术领域。该特异性识别猪ROSA26基因的sgRNA识别猪ROSA26基因的第一内含子，且用于CRISPR/Cpf1系统，含有如SEQ ID NO.1所示的负责识别靶片段的序列。该sgRNA配合CRISPR/Cpf1系统，敲除效率为27％且特异性强，在进行序列敲除时，能有效减少脱靶现象，减少非特异性切割对基因组带来的不利影响。