An endogenous mCherry-tagged COSA-1 as a crossover investigation tool in Caenorhabditis elegans.
An endogenous mCherry-tagged COSA-1 as a crossover investigation tool in Caenorhabditis elegans.
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DOI：
10.17912/micropub.biology.000627
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发表时间：
2022
影响因子：
--
通讯作者：
Wang, Bin
中科院分区：
文献类型：
--
作者：
Ezechukwu, Chiemekam Samuel;Odiba, Arome Solomon;Liao, Guiyan;Fang, Wenxia;Wang, Bin
梭线虫cosa-1基因编码交换位点相关蛋白1（COSA-1），这是一种细胞周期蛋白相关蛋白，在减数分裂期间促进交换（CO）。以往的研究表明，CO动力学在活C。线虫主要依赖于通过微粒轰击方法产生的绿色荧光蛋白标记的COSA-1转基因株。在此，我们将红色荧光蛋白mCherry插入到cosa-1基因的C末端，通过CRISPR/Cas9技术建立cosa-1：：mCherry转基因蠕虫。COSA-1：：mCherry在粗线期早期出现，在双线期消失，在粗线期晚期有6个COSA-1：：mCherry灶，与来自AV 630株的GFP：：COSA-1共定位。此外，cosa-1：：mCherry融合基因的转基因株系在窝卵数、子代存活率和雄性频率方面均无缺陷，为进一步研究家蚕减数分裂提供了有用的工具。优雅的
The C. elegans cosa-1 gene encodes the crossover site-associated-1 (COSA-1) protein, a cyclin-related protein that functions in promoting crossovers (COs) during meiosis. Previous studies regarding CO dynamics in live C. elegans have mostly relied on the green fluorescent protein-tagged cosa-1 transgenic strain, which was generated by the microparticle bombardment method. Here, we insert the red fluorescence protein mCherry at the C-terminal of the cosa-1 gene to establish cosa-1::mCherry transgenic worm by the CRISPR/Cas9 technique. The COSA-1::mCherry was observed to appear from the early pachytene, and disappear in the diplotene zone of the germline, with 6 COSA-1:: mCherry foci in the late pachytene, which colocalized with GFP::COSA-1 from AV630 strain. Furthermore, the transgenic strain harboring a cosa-1::mCherry fusion shows no defect in the brood size, progeny viability and male frequency, which provides a useful tool for the meiotic analysis in C. elegans .