Phenotypic Characterization of Parp-1 and Parp-2 Deficient Mice and Cells
Poly(ADP-ribosyl)ation is a post-translational modification of proteins mediated by Poly(ADP-ribose) polymerases (Parps), a family of 17 members. Among them, Poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 are so far the sole enzymes whose catalytic activity has been shown to be induced by DNA strand breaks. The generation and characterization of Parp-1 and Parp-2 deficient cellular and animal models have largely contributed to describe both proteins as active players of the base excision repair/single-strand break repair (BER/SSBR) process with both redundant and more specific functions. Double Parp-1−/−Parp-2−/− embryos die at gastrulation demonstrating the crucial role of poly(ADP-ribosyl)ation during embryonic development, whereas a specific female lethality related to X chromosome instability is associated with the Parp-1+/−Parp-2−/− genotype. Finally, recent research discovered emerging unique functions of Parp-2 in physiological processes including spermatogenesis, T-cell maturation, and adipogenesis although with distinct mechanisms. In this chapter, we describe standard operating procedures used to genotype and phenotype both mouse lines and the derived mouse embryonic fibroblasts.
Key wordsPoly(ADP-ribose) polymerases Genome integrity DNA damage Knockout mice Mouse embryonic fibroblasts
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