Single-Cell Sequencing Reveals Epigenetic Factors Involved in Senescence of Mouse Embryonic Fibroblasts (MEFs)

To enhance the efficiency of library infection, we generated Cre-Loxp expressing Cas9 mice to obtain MEF-Cas9 cells for replicative senescence studies. During the passaging of MEF-Cas9 primary cells, we observed a significant increase in both protein and mRNA levels of p16, as well as an increase in senescence-associated β-galactosidase staining, indicating that MEF cells stably expressing Cas9 can normally undergo replicative senescence with passaging. Subsequently, we infected MEF-Cas9 cells with a laboratory-constructed epigenetic factor gRNA library. After the third passage (P3) of primary cells being infected with the virus, we performed single-cell transcriptome sequencing on the seventh passage (P7) cells infected with the library and control cells .