Ezechukwu Chiemekam Samuel, Odiba Arome Solomon, Liao Guiyan, Fang Wenxia, Wang Bin
State Key Laboratory of Non-food Biomass and Enzyme Technology, Guangxi Academy of Sciences, Nanning 530007, China.
Department of Zoology and Environmental Biology, University of Nigeria, Nsukka 410001, Nigeria.
MicroPubl Biol. 2022 Aug 16;2022. doi: 10.17912/micropub.biology.000627. eCollection 2022.
The 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 have mostly relied on the green fluorescent protein-tagged transgenic strain, which was generated by the microparticle bombardment method. Here, we insert the red fluorescence protein mCherry at the C-terminal of the gene to establish 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 fusion shows no defect in the brood size, progeny viability and male frequency, which provides a useful tool for the meiotic analysis in .
该基因编码交叉位点相关蛋白1（COSA-1），这是一种细胞周期蛋白相关蛋白，在减数分裂过程中促进交叉（COs）。此前关于活体中CO动态的研究大多依赖于通过微粒轰击法产生的绿色荧光蛋白标记的转基因菌株。在这里，我们通过CRISPR/Cas9技术在该基因的C端插入红色荧光蛋白mCherry，以建立转基因线虫。观察到COSA-1::mCherry从粗线期早期出现，并在生殖系的双线期区域消失，在粗线期后期有6个COSA-1::mCherry焦点，与来自AV630菌株的GFP::COSA-1共定位。此外，携带融合体的转基因菌株在产卵量、后代活力和雄性频率方面没有缺陷，这为秀丽隐杆线虫的减数分裂分析提供了一个有用的工具。
