Abstract:
The unicellular green alga, Chlamydomonas reinhardtii, has played a central role in discovering much of what is currently known about the composition, assembly, and function of cilia and flagella. Chlamydomonas combines excellent genetics, such as the ability to grow cells as haploids or diploids and to perform tetrad analysis, with an unparalleled ability to detach and isolate flagella in a single step without cell lysis. The combination of genetics and biochemistry that is possible in Chlamydomonas has allowed many of the key components of the cilium to be identified by looking for proteins that are missing in a defined mutant. Few if any other model organisms allow such a seamless combination of genetic and biochemical approaches. Other major advantages of Chlamydomonas compared to other systems include the ability to induce flagella to regenerate in a highly synchronous manner, allowing the kinetics of flagellar growth to be measured, and the ability of Chlamydomonas flagella to adhere to glass coverslips allowing Intraflagellar Transport to be easily imaged inside the flagella of living cells, with quantitative precision and single-molecule resolution. These advantages continue to work in favor of Chlamydomonas as a model system going forward, and are now augmented by extensive genomic resources, a knockout strain collection, and efficient CRISPR gene editing. While Chlamydomonas has obvious limitations for studying ciliary functions related to animal development or organ physiology, when it comes to studying the fundamental biology of cilia and flagella, Chlamydomonas is simply unmatched in terms of speed, efficiency, cost, and the variety of approaches that can be brought to bear on a question.
摘要:
单细胞绿藻,莱茵衣藻,在发现许多目前已知的构图方面发挥了核心作用,装配,纤毛和鞭毛的功能。衣藻结合了优秀的遗传学,例如将细胞生长为单倍体或二倍体并进行四分体分析的能力,具有无与伦比的分离和分离鞭毛的能力,无需细胞裂解。在衣藻中可能的遗传学和生物化学的结合已经允许通过寻找在确定的突变体中缺失的蛋白质来鉴定纤毛的许多关键组分。几乎没有任何其他模型生物允许遗传和生化方法的这种无缝组合。与其他系统相比,衣藻的其他主要优点包括能够诱导鞭毛以高度同步的方式再生,可以测量鞭毛生长的动力学,以及衣藻鞭毛粘附在玻璃盖玻片上的能力,允许在活细胞鞭毛内轻松成像,具有定量精度和单分子分辨率。这些优势继续有利于衣藻作为一个模型系统向前发展,现在被广泛的基因组资源所增强,敲除菌株集合,和高效的CRISPR基因编辑。虽然衣藻在研究与动物发育或器官生理相关的纤毛功能方面有明显的局限性,当谈到研究纤毛和鞭毛的基本生物学时,就速度而言,衣藻是无与伦比的,效率,成本,以及可以解决一个问题的各种方法。