Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
BMC Genet. 2010 Nov 30;11:106. doi: 10.1186/1471-2156-11-106.
Forward genetic screens in mice provide an unbiased means to identify genes and other functional genetic elements in the genome. Previously, a large scale ENU mutagenesis screen was conducted to query the functional content of a ~50 Mb region of the mouse genome on proximal Chr 5. The majority of phenotypic mutants recovered were embryonic lethals.
We report the high resolution genetic mapping, complementation analyses, and positional cloning of mutations in the target region. The collection of identified alleles include several with known or presumed functions for which no mutant models have been reported (Tbc1d14, Nol14, Tyms, Cad, Fbxl5, Haus3), and mutations in genes we or others previously reported (Tapt1, Rest, Ugdh, Paxip1, Hmx1, Otoe, Nsun7). We also confirmed the causative nature of a homeotic mutation with a targeted allele, mapped a lethal mutation to a large gene desert, and localized a spermiogenesis mutation to a region in which no annotated genes have coding mutations. The mutation in Tbc1d14 provides the first implication of a critical developmental role for RAB-GAP-mediated protein transport in early embryogenesis.
This collection of alleles contributes to the goal of assigning biological functions to all known genes, as well as identifying novel functional elements that would be missed by reverse genetic approaches.
在小鼠中进行正向遗传学筛选为鉴定基因组中的基因和其他功能遗传元件提供了一种无偏的方法。此前，进行了一项大规模的ENU 诱变筛选，以查询小鼠基因组近端 Chr5 上约 50Mb 区域的功能内容。回收的大多数表型突变体都是胚胎致死的。
我们报告了目标区域突变的高分辨率遗传作图、互补分析和定位克隆。鉴定的等位基因集合包括几个具有已知或假定功能的等位基因，但尚未报道其突变模型（Tbc1d14、Nol14、Tyms、Cad、Fbxl5、Haus3），以及我们或其他人之前报道过的基因中的突变（Tapt1、Rest、Ugdh、Paxip1、Hmx1、Otoe、Nsun7）。我们还证实了一个靶向等位基因的同源突变的因果性质，将一个致死突变映射到一个大的基因荒漠中，并将一个精子发生突变定位到一个没有注释基因具有编码突变的区域。Tbc1d14 中的突变首次表明 RAB-GAP 介导的蛋白质运输在早期胚胎发生中具有关键的发育作用。
该等位基因集合有助于将所有已知基因的生物学功能分配的目标，并鉴定通过反向遗传学方法可能错过的新的功能元件。
