Gene Editing in Trypanosomatids: Tips and Tricks in the CRISPR-Cas9 Era

Akila Yagoubat (1) , Rosa Corrales (1) , Patrick Bastien (2, 3) , Maude Lévêque (2, 3) , Yvon Sterkers (2, 3)

1 MIVEGEC - Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle
2 MIVEGEC-BioGEPPE - Biologie, Génétique et Pathologie des Pathogènes Eucaryotes
3 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]

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.

Mots clés: CRE-Lox; CRISPR-Cas9; Leishmania; Trypanosoma; UTR modification; inducible genome editing

Domaines: Génétique

Akila Yagoubat, Rosa Corrales, Patrick Bastien, Maude Lévêque, Yvon Sterkers. Gene Editing in Trypanosomatids: Tips and Tricks in the CRISPR-Cas9 Era. Trends in Parasitology, 2020, 36 (9), pp.745-760. ⟨10.1016/j.pt.2020.06.005⟩. ⟨hal-02920067⟩