Abstract: Objective：To construct the Msx1-Cre mouse model. Methods：Design two sgRNA in the CRISPR/Cas9 system that target the Msx1 gene and construct homologous recombination fragments containing Cre sequences; Construct sgRNA vector plasmids and verify the target efficiency and off-target situation of sgRNA at the cellular level; Obtain F0 generation mice knocked into Cre sequence of Msx1 gene by microinjection method; Identification and functional verification of mouse models. Results：The designed sgRNA effectively cut the target site downstream of the Msx1 promoter in a targeted manner; We successfully obtained the Msx1Cre mouse model by microinjection; GFP was specifically located on the membrane of Msx1+ cells in Msx1Cre;R26RmTmG mice. Conclusions：The Msx1Cre mouse model was successfully constructed based on the CRISPR/Cas9 system.

Key words: CRISPR/Cas9, Msx1, Cre, mouse models