Shah, R.; van den Berk, P.C.M.; Pritchard, C.E.J.; Song, J.-Y.; Kreft, M.; Pilzecker, B.; Jacobs, H. A C57BL/6J Fancg-KO Mouse Model Generated by CRISPR/Cas9 Partially Captures the Human Phenotype. Int. J. Mol. Sci.2023, 24, 11129.
Shah, R.; van den Berk, P.C.M.; Pritchard, C.E.J.; Song, J.-Y.; Kreft, M.; Pilzecker, B.; Jacobs, H. A C57BL/6J Fancg-KO Mouse Model Generated by CRISPR/Cas9 Partially Captures the Human Phenotype. Int. J. Mol. Sci. 2023, 24, 11129.
Shah, R.; van den Berk, P.C.M.; Pritchard, C.E.J.; Song, J.-Y.; Kreft, M.; Pilzecker, B.; Jacobs, H. A C57BL/6J Fancg-KO Mouse Model Generated by CRISPR/Cas9 Partially Captures the Human Phenotype. Int. J. Mol. Sci.2023, 24, 11129.
Shah, R.; van den Berk, P.C.M.; Pritchard, C.E.J.; Song, J.-Y.; Kreft, M.; Pilzecker, B.; Jacobs, H. A C57BL/6J Fancg-KO Mouse Model Generated by CRISPR/Cas9 Partially Captures the Human Phenotype. Int. J. Mol. Sci. 2023, 24, 11129.
Abstract
Fanconi anemia (FA) develops due to a mutation in one of the FANC genes that are involved in repair of interstrand crosslinks (ICLs). FANCG, a member of the FA core complex, is essential for ICL repair. Previous FANCG-deficient mouse models were generated with drug-based selection cassettes in mixed mice backgrounds, leading to a disparity in interpretation of genotype-related phenotype. To exclude these confounders, we created a Fancg-KO (KO) mouse model using CRISPR/Cas9. The entire Fancg locus was targeted and maintained on the immunological well-characterized C57BL/6J background. Intercrossing of heterozygous mice resulted in sub-Mendelian numbers of homozygous mice, suggesting loss of FANCG can be embryonic lethal. KO mice displayed infertility, hypogonadism but no other developmental problems. Bone marrow analysis revealed a defect in various hematopoietic stem and progenitor subsets with a bias towards myelopoiesis. Cell lines derived from Fancg-KO mice were hypersensitive to crosslinking agents-cisplatin and Mitomycin C, and Fancg-KO mouse embryonic fibroblasts (MEFs) displayed increased γ-H2AX upon cisplatin treatment. Reconstitution of these MEFs with Fancg cDNA corrected for the ICL hypersensitivity. Collectively, this project provides a new, genetically and immunologically well-defined Fancg-KO mouse model for further in vivo and in vitro studies on FANCG and ICL repair.
Keywords
Fanconi anemia (FA); FANCG, interstrand crosslink (ICL); DNA damage response; genetically engineered mouse model (GEMM); CRISPR/Cas9; cisplatin; Mitomycin C (MMC); hematopoiesis
Subject
Biology and Life Sciences, Life Sciences
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
