The MSLN gene encodes a preproprotein that is proteolytically processed into two distinct proteins: megakaryocyte potentiating factor (MPF) and mesothelin ^[1]. MPF functions as a cytokine that promotes the formation of megakaryocytes in bone marrow, while mesothelin is a glycosylphosphatidylinositol-anchored cell-surface protein that may play a role in cell adhesion. While mesothelin expression is normally low and restricted to mesothelial cells lining the pleura, peritoneum, and pericardium, it is highly overexpressed in several epithelial cancers, including malignant mesothelioma, ovarian cancer, pancreatic cancer, and certain types of lung cancer ^[2-3]. This high expression and its function in promoting cell proliferation and metastasis make mesothelin a significant biomarker and a target for cancer therapies ^[4]. The B6-huMSLN mouse model was generated by replacing sequences from the ATG start codon to the TGA stop codon of the endogenous mouse Msln gene with the sequences from the ATG start codon to the TGA stop codon of the human MSLN gene. This model can be used to study the pathological mechanisms and therapeutic approaches for several cancers, including malignant mesothelioma, ovarian cancer, pancreatic cancer, and certain types of lung cancer, as well as for the development of MSLN-targeted drugs.

Keap1 is located on chromosome 9 of mice. Nuclease Technology was used to design sgRNA; Keap1 knockout mice were obtained by applying high-throughput electroporation of fertilized eggs. After sexual maturity, sperm were collected for cryopreservation.

Slc46a2 is located on chromosome 4 of mice. SgRNA and ssDNA will be designed using Nuclease Technology; Slc46a2 conditional knockout mice will be obtained by high-throughput electroporation of fertilized eggs. After sexual maturity, sperm will be collected for cryopreservation.