CRISPR/CAS9 SYSTEM AND APPLICATION THEREOF IN CONSTRUCTION OF α, β, AND αβ THALASSEMIA MODEL PORCINE CELL LINES

摘要

A CRISPR/Cas9 system and an application thereof in the construction of α, β, and α&β thalassemia model porcine cell lines. A pair of target sequences are designed for each of porcine HBA and HBB genes. Three types of CRISPR/Cas9 systems are constructed using the two pairs of target sequences. Each type of CRISPR/Cas9 system comprises a gRNA vector and a Cas9 expression vector. The expression vectors in the three types of CRISPR/Cas9 systems are respectively transferred into porcine fibroblasts in proportion, and are screened to obtain α, β, and α&β thalassemia model porcine cell lines. Single gene mutations and combined gene mutations are respectively caused to porcine HBA and HBB genes in porcine primary fibroblasts by means of a gene editing technology, to obtain HBA, HBB, and HBA&HBB gene-mutated cells.