Recent advances in genome editing of bloodstream forms of Trypanosoma congolense using CRISPR-Cas9 ribonucleoproteins: Proof of concept

Minet Cécile, Chantal Isabelle, Berthier David. 2023. Recent advances in genome editing of bloodstream forms of Trypanosoma congolense using CRISPR-Cas9 ribonucleoproteins: Proof of concept. Experimental Parasitology, 252:108589, 10 p.

Résumé : African Animal Trypanosomosis (AAT or Nagana) is a vector-borne disease caused by Trypanosomatidae, genus Trypanosoma. The disease is transmitted by the bite of infected hematophagous insects, mainly tsetse flies but also other blood-sucking insects including stomoxes and tabanids. Although many trypanosome species infect animals, the main agents responsible for this disease with a strong socio-economic and veterinary health impact are Trypanosoma congolense (T. congolense or Tc), Trypanosoma vivax (T.vivax), and to a lesser extent, Trypanosoma brucei brucei (T.brucei brucei or Tbb). These parasites mainly infect livestock, including cattle, in sub-Saharan Africa, with major repercussions in terms of animal productivity and poverty for populations which are often already very poor. As there is currently no vaccine, the fight against the disease is primarily based on diagnosis, treatment and vector control. To develop new tools (particularly therapeutic tools) to fight against the disease, we need to know both the biology and the genes involved in the pathogenicity and virulence of the parasites. To date, unlike for Trypanosoma brucei (T.brucei) or Trypanosoma cruzi (T.cruzi), genome editing tools has been relatively little used to study T. congolense. We present an efficient, reproducible and stable CRISPR-Cas9 genome editing system for use in Tc bloodstream forms (Tc-BSF). This plasmid-free system is based on transient expression of Cas9 protein and the use of a ribonucleoprotein formed by the Cas9 and sgRNA complex. This is the first proof of concept of genome editing using CRISPR-Cas9 ribonucleoproteins on Tc-BSF. This adapted protocol enriches the “toolbox” for the functional study of genes of interest in blood forms of the Trypanosoma congolense. This proof of concept is an important step for the scientific community working on the study of trypanosomes and opens up new perspectives for the control of and fight against animal trypanosomosis.

Mots-clés Agrovoc : Trypanosoma congolense, trypanosomose, Trypanosoma, Trypanosoma vivax, Trypanosoma brucei, maladie transmise par vecteur, contrôle de maladies, maladie des animaux, Trypanosoma cruzi, vecteur de maladie, trypanosomose africaine, génome, diagnostic, bovin, épidémiologie

Mots-clés libres : Trypanosoma congolense bloodstream forms (Tc- BSF), Transfection Ribonucleoproteins, CRISPR Genome editing, Genome editing, Ribonucleopropteins

Classification Agris : L73 - Maladies des animaux

Champ stratégique Cirad : CTS 4 (2019-) - Santé des plantes, des animaux et des écosystèmes

Agences de financement hors UE : Agence Nationale de la Recherche, Centre de Coopération Internationale en Recherche Agronomique pour le Développement, Alliance Française contre les Maladies Parasitaires

Projets sur financement : (FRA) Développment d'une infrastructure française distribuée coordonnée, (FRA) Alliance française contre les maladies parasitaires

Auteurs et affiliations

• Minet Cécile, CIRAD-BIOS-UMR INTERTRYP (FRA) ORCID: 0000-0002-9287-1481 - auteur correspondant
• Chantal Isabelle, CIRAD-BIOS-UMR INTERTRYP (FRA) ORCID: 0000-0002-3429-484X
• Berthier David, CIRAD-BIOS-UMR INTERTRYP (FRA) ORCID: 0000-0002-3283-6588

Source : Cirad-Agritrop (https://agritrop.cirad.fr/605730/)

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