In the latest issue of the journal Cell, back-to-back manuscripts are published detailing the application of CRISPR base editors to interrogate the functional significance of nucleotide variants in genes that operate in the DNA damage response (DDR). In the lab of John Doench, initial screens involved nucleotide variants in the cancer predisposition genes BRCA1/2 and anti-apoptotic genes MCL1 and BCL2L1. Furthermore, massive pooled screens were performed targeting over 3500 genes with the potential to test the functional significance of over 52k nucleotide variants reported in the ClinVar database. The group of Alberto Ciccia also used cytosine base editors with over 2k sgRNAs in order to create nucleotide variants in 86 genes involved in DNA replication, damage signaling, homology directed and interstrand crosslink repair. The generated data provide new insights into functional protein domains and identified nucleotide variants associated with gain or loss of function in numerous DDR genes.

For more information, see:
Hanna, R.E., et al. (2021) Massively parallel assessment of human variants with base editor screens. Cell 184: 1064-1080.e20. https://doi.org/10.1016/j.cell.2021.01.012.
Cuella-Martin, R., et al. (2021) Functional interrogation of DNA damage response variants with base editing screens. Cell 184: 1081-1097.e19. https://doi.org/10.1016/j.cell.2021.01.041.
Koch, L. (2021) Finding function with base editing screens. Nature Reviews Genetics https://doi.org/10.1038/s41576-021-00340-0.

Keywords: CRISPR, base editors, gRNA tiling, screens, variants of uncertain significance (VUS), DDR

Questions? Email: crispr@amsterdamumc.nl