Department of Genome Sciences, University of Washington, Seattle, Washington 98195-5065, USA.
Genetics. 2010 Mar;184(3):853-63. doi: 10.1534/genetics.109.110270. Epub 2009 Dec 14.
Comparative studies of Caenorhabditis briggsae and C. elegans have provided insights into gene function and developmental control in both organisms. C. elegans is a well developed model organism with a variety of molecular and genetic tools to study gene functions. In contrast, there are only very limited tools available for its closest relative, C. briggsae. To take advantage of the full potential of this comparative approach, we have developed several genetic and molecular tools to facilitate functional analysis in C. briggsae. First, we designed and implemented an SNP-based oligonucleotide microarray for rapid mapping of genetic mutants in C. briggsae. Second, we generated a mutagenized frozen library to permit the isolation of targeted deletions and used the library to recover a deletion mutant of cbr-unc-119 for use as a transgenic marker. Third, we used the cbr-unc-119 mutant in ballistic transformation and generated fluorescently labeled strains that allow automated lineaging and cellular resolution expression analysis. Finally, we demonstrated the potential of automated lineaging by profiling expression of egl-5, hlh-1, and pha-4 at cellular resolution and by detailed phenotyping of the perturbations on the Wnt signaling pathway. These additions to the experimental toolkit for C. briggsae should greatly increase its utility in comparative studies with C. elegans. With the emerging sequence of nematode species more closely related to C. briggsae, these tools may open novel avenues of experimentation in C. briggsae itself.
秀丽隐杆线虫和 C. briggsae 的比较研究为这两种生物的基因功能和发育调控提供了深入的了解。秀丽隐杆线虫是一种发育良好的模式生物，具有多种分子和遗传工具来研究基因功能。相比之下，它最接近的亲属 C. briggsae 仅有非常有限的工具可用。为了充分利用这种比较方法的潜力，我们开发了几种遗传和分子工具，以促进 C. briggsae 中的功能分析。首先，我们设计并实施了基于 SNP 的寡核苷酸微阵列，用于快速绘制 C. briggsae 中的遗传突变体图谱。其次，我们生成了诱变冷冻文库，以允许分离靶向缺失，并使用该文库回收 cbr-unc-119 的缺失突变体，用作转基因标记。第三，我们使用 cbr-unc-119 突变体进行弹道转化，并生成荧光标记的菌株，允许自动谱系分析和细胞分辨率表达分析。最后，我们通过在细胞分辨率下对 egl-5、hlh-1 和 pha-4 的表达进行自动谱系分析，并通过对 Wnt 信号通路的干扰进行详细表型分析，证明了自动谱系分析的潜力。这些 C. briggsae 实验工具包的增加应该会大大提高其在与秀丽隐杆线虫比较研究中的实用性。随着与 C. briggsae 关系更密切的线虫物种序列的出现，这些工具可能会为 C. briggsae 本身开辟新的实验途径。
