A C57BL/6J Fancg-KO Mouse Model Generated by CRISPR/Cas9 Partially Captures the Human Phenotype

Fanconi anemia (FA) develops due to a mutation in one of the FANC genes that are involved in the 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 m...

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Vydáno v:International journal of molecular sciences Ročník 24; číslo 13; s. 11129
Hlavní autoři: Shah, Ronak, van den Berk, Paul C. M., Pritchard, Colin E. J., Song, Ji-Ying, Kreft, Maaike, Pilzecker, Bas, Jacobs, Heinz
Médium: Journal Article
Jazyk:angličtina
Vydáno: Switzerland MDPI AG 01.07.2023
MDPI
Témata:
Ablation
Analysis
Anemia
Animals
Bone marrow
Cisplatin - metabolism
CRISPR
CRISPR-Cas Systems
Crosslinked polymers
DNA-Binding Proteins - metabolism
Fanconi Anemia - genetics
Fanconi Anemia - metabolism
Fanconi Anemia Complementation Group G Protein - genetics
Fanconi's anemia
Fibroblasts
Fibroblasts - metabolism
Genetic aspects
Genotype & phenotype
Humans
Immunotherapy
Infertility
Life expectancy
Metabolism
Mice
Mice, Inbred C57BL
Mitomycin
Phenotype
Proteins
Stem cells
Thymus gland
Netherlands
Germany
fanconi anemia (FA)
cisplatin (CsPt)
hematopoiesis
Mitomycin C (MMC)
CRISPR/Cas9
DNA damage response
interstrand crosslink (ICL)
genetically engineered mouse model (GEMM)
FANCG
ISSN:1422-0067, 1661-6596, 1422-0067
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Popis
Shrnutí:Fanconi anemia (FA) develops due to a mutation in one of the FANC genes that are involved in the 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 the interpretation of genotype-related phenotype. We created a Fancg-KO (KO) mouse model using CRISPR/Cas9 to exclude these confounders. The entire Fancg locus was targeted and maintained on the immunological well-characterized C57BL/6J background. The intercrossing of heterozygous mice resulted in sub-Mendelian numbers of homozygous mice, suggesting the loss of FANCG can be embryonically lethal. KO mice displayed infertility and 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 the crosslinking agents cisplatin and Mitomycin C, and Fancg-KO mouse embryonic fibroblasts (MEFs) displayed increased γ-H2AX upon cisplatin treatment. The reconstitution of these MEFs with Fancg cDNA corrected for the ICL hypersensitivity. 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.
Bibliografie:ObjectType-Article-1
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ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms241311129