CRISPR/Cas9 technology has revolutionized biology. This prokaryotic defense system against foreign DNA has been repurposed for genome editing in a broad range of cell tissues and organisms. Trypanosomatids are flagellated protozoa belonging to the order Kinetoplastida. Some of its most representative members cause important human diseases affecting millions of people worldwide, such as Chagas disease, sleeping sickness and different forms of leishmaniases. Trypanosomatid infections represent an enormous burden for public health and there are no effective treatments for most of the diseases they cause. Since the emergence of the CRISPR/Cas9 technology, the genetic manipulation of these parasites has notably improved. As a consequence, genome editing is now playing a key role in the functional study of proteins, in the characterization of metabolic pathways, in the validation of alternative targets for antiparasitic interventions, and in the study of parasite biology and pathogenesis. In this work we review the different strategies that have been used to adapt the CRISPR/Cas9 system to Trypanosoma cruzi, Trypanosoma brucei, and Leishmania spp., as well as the research progress achieved using these approaches. Thereby, we will present the state‐of‐the‐art molecular tools available for genome editing in trypanosomatids to finally point out the future perspectives in the field.

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用于锥虫基因组编辑的最先进 CRISPR/Cas9 技术

CRISPR/Cas9 技术彻底改变了生物学。这种针对外来 DNA 的原核防御系统已被重新用于各种细胞组织和生物体的基因组编辑。锥虫是属于动质体目（Kinetoplastida）的鞭毛原生动物。它的一些最具代表性的成员会导致影响全世界数百万人的重要人类疾病，例如恰加斯病、昏睡病和不同形式的利什曼病。锥虫感染对公共健康构成了巨大的负担，并且对它们引起的大多数疾病没有有效的治疗方法。自从 CRISPR/Cas9 技术出现以来，对这些寄生虫的基因操作有了显着改善。因此，基因组编辑现在在蛋白质的功能研究中发挥着关键作用，在代谢途径的表征，验证抗寄生虫干预的替代目标以及寄生虫生物学和发病机制的研究中。在这项工作中，我们回顾了用于使 CRISPR/Cas9 系统适应克氏锥虫、布氏锥虫和利什曼原虫的不同策略，以及使用这些方法取得的研究进展。因此，我们将展示可用于锥虫基因组编辑的最先进的分子工具，以最终指出该领域的未来前景。和利什曼原虫，以及使用这些方法取得的研究进展。因此，我们将展示可用于锥虫基因组编辑的最先进的分子工具，以最终指出该领域的未来前景。和利什曼原虫，以及使用这些方法取得的研究进展。因此，我们将展示可用于锥虫基因组编辑的最先进的分子工具，以最终指出该领域的未来前景。