Gene editing in trypanosomatids has long been proven difficult. The development of CRISPR-Cas9 has improved this issue, opening the way to a better understanding of biological processes and drug-resistance mechanisms, and screening of drug targets. Different strategies have now been developed: either PCR- or plasmid-based, differing mainly in the nature of the donor DNA and the single guide RNA transcription. Here we review the main genetic tools available for Leishmania spp., Trypanosoma cruzi, and Trypanosoma brucei for gene tagging, single-base editing, and deletion of nonessential and essential genes. We discuss the main advantages and challenges of different strategies and how to choose 'the right cut' depending on the importance of untranslated regions. These considerations allow selection of the most accurate gene editing approach for a given functional analysis.

长期以来，锥虫的基因编辑已被证明是困难的。CRISPR-Cas9的开发改善了这个问题，为更好地了解生物学过程和耐药机制以及筛选药物靶标开辟了道路。现在已经开发出不同的策略：基于PCR的或基于质粒的策略，主要区别在于供体DNA和单个指导RNA转录的性质。在这里，我们回顾了可用于利什曼原虫属，锥虫锥虫和布鲁氏锥虫的主要遗传工具。用于基因标记，单碱基编辑以及非必需和必需基因的删除。我们讨论了不同策略的主要优势和挑战，以及如何根据未翻译区域的重要性选择“正确切入”。这些考虑因素允许针对给定的功能分析选择最准确的基因编辑方法。