Abstract
Maintenance of genome integrity via repair of DNA damage is a key biological process required to suppress diseases, including Fanconi anemia (FA). We generated loss-of-function human haploid cells for FA complementation group C (FANCC), a gene encoding a component of the FA core complex, and used genome-wide CRISPR libraries as well as insertional mutagenesis to identify synthetic viable (genetic suppressor) interactions for FA. Here we show that loss of the BLM helicase complex suppresses FANCC phenotypes and we confirm this interaction in cells deficient for FA complementation group I and D2 (FANCI and FANCD2) that function as part of the FA I-D2 complex, indicating that this interaction is not limited to the FA core complex, hence demonstrating that systematic genome-wide screening approaches can be used to reveal genetic viable interactions for DNA repair defects.
Highlights
Maintenance of genome integrity via repair of DNA damage is a key biological process required to suppress diseases, including Fanconi anemia (FA)
To identify synthetic viable interactions for FANCC, we set up two genome-wide approaches to screen for mutations that alleviate the hypersensitivity of ΔFANCC cells to mitomycin C (MMC)-induced DNA damage (Fig. 1a)
By comparing enriched genes in the CRISPR screen performed on MMC-treated ΔFANCC cells to enriched genes identified by an additional CRISPR screen performed on MMC-treated WT cells, we found that loss of the BLM complex rescued ΔFANCC but had little or no effect in WT cells (Supplementary Fig. 4a)
Summary
Maintenance of genome integrity via repair of DNA damage is a key biological process required to suppress diseases, including Fanconi anemia (FA). We generated loss-of-function human haploid cells for FA complementation group C (FANCC), a gene encoding a component of the FA core complex, and used genome-wide CRISPR libraries as well as insertional mutagenesis to identify synthetic viable (genetic suppressor) interactions for FA. We identified synthetic viable interactions for FA by performing genome-wide screens on isogenic human haploid cells lacking the FA complementation group C (FANCC) protein, following exposure to the DNA ICL-inducing agent mitomycin C (MMC). We identify the BLM helicase complex as a suppressor of Fanconi anemia phenotypes in human cells, demonstrating that systematic screening approaches can be used to reveal genetic viable interactions for DNA repair defects
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