Lee Chang-Hyun, Rimesso Gerard, Reynolds David M, Cai Jinlu, Baker Nicholas E
Department of Genetics, Albert Einstein College of Medicine, Bronx, New York 10461.
Department of Genetics, Albert Einstein College of Medicine, Bronx, New York 10461
G3 (Bethesda). 2016 Oct 13;6(10):3207-3217. doi: 10.1534/g3.116.029421.
Cell competition, the conditional loss of viable genotypes only when surrounded by other cells, is a phenomenon observed in certain genetic mosaic conditions. We conducted a chemical mutagenesis and screen to recover new mutations that affect cell competition between wild-type and RpS3 heterozygous cells. Mutations were identified by whole-genome sequencing, making use of software tools that greatly facilitate the distinction between newly induced mutations and other sources of apparent sequence polymorphism, thereby reducing false-positive and false-negative identification rates. In addition, we utilized iPLEX MassARRAY for genotyping recombinant chromosomes. These approaches permitted the mapping of a new mutation affecting cell competition when only a single allele existed, with a phenotype assessed only in genetic mosaics, without the benefit of complementation with existing mutations, deletions, or duplications. These techniques expand the utility of chemical mutagenesis and whole-genome sequencing for mutant identification. We discuss mutations in the Atm and Xrp1 genes identified in this screen.
细胞竞争是指在某些遗传嵌合条件下，仅当被其他细胞包围时，存活基因型的条件性丧失现象。我们进行了化学诱变和筛选，以恢复影响野生型和RpS3杂合细胞之间细胞竞争的新突变。通过全基因组测序鉴定突变，利用软件工具极大地促进了新诱导突变与其他明显序列多态性来源之间的区分，从而降低了假阳性和假阴性识别率。此外，我们利用iPLEX MassARRAY对重组染色体进行基因分型。这些方法允许在仅存在单个等位基因时定位影响细胞竞争的新突变，其表型仅在遗传嵌合体中评估，而无需与现有突变、缺失或重复进行互补。这些技术扩展了化学诱变和全基因组测序在突变体鉴定中的应用。我们讨论了在此筛选中鉴定出的Atm和Xrp1基因中的突变。
