Construction of gene editing system of central precocious puberty model pig nuclear transfer donor cells with mkrn3 gene mutation and application thereof

Abstract

The application discloses a gene editing system for constructing a central congenital sexual precocity model pig nuclear transfer donor cell of an MKRN3 gene mutation and application thereof. The application provides a method for preparing a recombinant cell, and the method comprises the following steps: co-transfecting MKRN3-gRNA3 shown in SEQ ID NO: 16, MKRN3-gRNA4 shown in SEQ ID NO: 17 and NCN protein into a pig cell to obtain the recombinant cell. The application adopts CRISPR / Cas9 technology combined with double gRNA editing to knockout the MKRN3 gene, simulates natural pathogenic genetic characteristics of central congenital sexual precocity and central congenital sexual precocity combined with lung cancer, and obtains a single cell clone of the MKRN3 gene knockout, thereby laying a foundation for cultivating a model pig of central congenital sexual precocity and central congenital sexual precocity combined with lung cancer through somatic cell nuclear transfer animal cloning technology in the later stage. The application has great application value for the research and development of drugs for central congenital sexual precocity and central congenital sexual precocity combined with lung cancer and the revelation of the pathogenesis of the diseases.

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