Jinyu Fu, Yunyi Gao, Feng Gao, Yalan Sheng, Saleh A Al-Farraj, Zigui Chen, Chundi Wang
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引用次数: 0
摘要
微生物拓展了我们对生物多样性的认识,并展现出重要的研究价值。纤毛原生动物是真核微生物中的一个高度分化的群体,具有许多纤毛、二形核和全基因组基因重排等特殊特征,是揭示许多生物过程的理想研究模型。对纤毛虫嗜热四膜虫的研究极大地促进了我们对 RNA 自拼接和酶活性、端粒生物学以及诺贝尔奖得主端粒酶机制的了解。纤毛虫有性生殖(结合)过程中的基因组重排涉及约 25-90% 的种系 DNA 的消除,这为研究大规模基因组重塑提供了机会,同时也揭示了一种更彻底的转座子抑制模式,即体细胞核中 Piwi-interacting 小 RNA 介导的 DNA 删除。虽然已经取得了很大进展,但研究主要集中在少数适合基因编辑的模式物种上。对于其他物种,虽然它们更适合解决一些科学空白,但由于基因工程的限制,研究无法开展。在此,我们总结了现有的纤毛虫基因工程策略,期望能为纤毛虫和其他尚不能进行基因编辑的生物提供基因工程工具的开发和优化灵感。
An overview of genetic manipulation tools in the studies of ciliates, with emphasis on gene knockout, knockdown, and overexpression
Microorganisms have expanded our understanding of biodiversity and exhibit crucial research values. Ciliated protists are a highly differentiated group of eukaryotic microorganisms with exceptional features, such as numerous cilia, dimorphic nuclei, and genome-wide gene rearrangement, that have made them ideal research models for revealing many biological processes. Studies of the ciliate Tetrahymena thermophila have greatly advanced our understanding of RNA self-splicing and enzymatic activity, telomere biology, and the Nobel Prize-winning telomerase mechanisms. Genome rearrangement during the sexual reproduction (conjugation) of ciliates, involving the elimination of ~25–90% of germline DNA, provides an opportunity to study large-scale genome remodelling while also revealing a more thorough mode of transposon repression, i.e. Piwi-interacting small RNA-mediated DNA deletion in the somatic nucleus. Although much progress has been made, research has focused mainly on a few model species that are amenable to gene editing. For other species, although they are more suitable to address some scientific gaps, research cannot be carried out owing to limitations of genetic engineering. Here, we summarize the existing genetic engineering strategies for ciliates, expecting to provide inspiration for the development and optimization of genetic engineering tools for ciliates and other organisms cannot yet be edited genetically.