Identification of a molecular network of DNA damage-induced neural cell death. Mus musculus

DNA damage plays a major role in neural cell death by necrosis and/or apoptosis. However, our understanding of the molecular mechanisms of neural cell death remains still incomplete. To acquire a global understanding of the various mediators related to DNA damage-induced neural cell death pathways, we performed a whole genomic wide screen in neural stem cells by using a siRNA library. We identified 80 genes required for DNA damage-induced cell death. 14 genes (17.5%) are directly related to cell death and/or apoptosis. 66 genes have not been previously directly linked to DNA damage-induced cell death. Using an integrated approach with functional and bioinformatics analysis, we have uncovered a molecular network containing several partially overlapping and interconnected pathways and/or protein complexes that are required for DNA damage-induced neural cell death. The identification of the network of neural cell death mediators will greatly enhance our understanding of the molecular mechanisms of neural cell death and provide therapeutic targets for nervous system disorders. Overall design: The note that new synthesized or activated cell death-inducing proteins are required for DNA damage-induced cell death leads us to propose that cells would be rescued from DNA damage-induced cell death if the cell death-inducing factors were knockdown by siRNAs. To obtain a global understanding of the network of molecular signaling pathways related to DNA damage-induced neural cell death, we performed a whole genome-wide screening using mouse 40k FIV lentiviral small interfering RNA (siRNA) library. We used PCR to amplify siRNA inserts from surviving cells after MNNG (N-methyl-N′-nitro-N-nitrosoguanidine) treatment and reference cells without treatment. The PCR products were labeled with biotin and applied to hybridize with the Affymetrix GeneChip® Mouse Genome 430 2.0 Array to identify surviving cell-associated siRNAs.