Converting Blastocrithidia Nonstop, a Trypanosomatid With Non-Canonical Genetic Code, Into a Genetically-Tractable Model

IF 2.6 2区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Microbiology Pub Date : 2025-04-09 DOI:10.1111/mmi.15365

Arnau Galan, Natalya Kraeva, Kristína Záhonová, Anzhelika Butenko, Alexei Yu Kostygov, Zdeněk Paris, Jiří Pergner, Claretta Bianchi, Fadel Fakih, Andreu Saura, Julius Lukeš, Vyacheslav Yurchenko

引用次数: 0

Abstract

Blastocrithidia nonstop is a protist with a highly unusual nuclear genetic code, in which all three standard stop codons are reassigned to encode amino acids, with UAA also serving as a sole termination codon. In this study, we demonstrate that this parasitic flagellate is amenable to genetic manipulation, enabling gene ablation and protein tagging. Using preassembled Cas9 ribonucleoprotein complexes, we successfully disrupted and tagged the non-essential gene encoding catalase. These advances establish this single-celled eukaryote as a model organism for investigating the malleability and evolution of the genetic code in eukaryotes.

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将非规范遗传密码的锥虫Blastocrithidia Nonstop转化为遗传可处理模型

胚核不间断是一种具有非常不寻常的核遗传密码的原生生物，其中所有三个标准终止密码子都被重新分配用于编码氨基酸，其中UAA也作为唯一的终止密码子。在这项研究中，我们证明了这种寄生鞭毛虫是适合基因操作的，使基因消融和蛋白质标记成为可能。利用预组装的Cas9核糖核蛋白复合物，我们成功地破坏并标记了编码过氧化氢酶的非必需基因。这些进展确立了这种单细胞真核生物作为研究真核生物遗传密码的延展性和进化的模式生物。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Microbiology 生物-生化与分子生物学

CiteScore

7.20

自引率

5.60%

发文量

132

审稿时长

1.7 months

期刊介绍： Molecular Microbiology, the leading primary journal in the microbial sciences, publishes molecular studies of Bacteria, Archaea, eukaryotic microorganisms, and their viruses. Research papers should lead to a deeper understanding of the molecular principles underlying basic physiological processes or mechanisms. Appropriate topics include gene expression and regulation, pathogenicity and virulence, physiology and metabolism, synthesis of macromolecules (proteins, nucleic acids, lipids, polysaccharides, etc), cell biology and subcellular organization, membrane biogenesis and function, traffic and transport, cell-cell communication and signalling pathways, evolution and gene transfer. Articles focused on host responses (cellular or immunological) to pathogens or on microbial ecology should be directed to our sister journals Cellular Microbiology and Environmental Microbiology, respectively.