Résumé
African Animal Trypanosomosis is a major vector-borne disease affecting livestock breading in Africa. It is caused by trypanosomes, especially Trypanosoma congolense with major negative impacts on animal production and negative repercussions in terms of poverty. To develop new tools to fight against the disease, we need to improve knowledge of the genes involved in the pathogenicity and virulence of the parasites as well as their functions. To date, unlike for Trypanosoma brucei or Trypanosoma cruzi, genome editing tools have been used relatively little to study T. congolense. We present an efficient, reproducible and stable CRISPR-Cas9 genome editing system for use in T. congolense 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 in Tc-BSF. This adapted protocol enriches the "toolbox" for the functional study of genes of interest in bloodstream forms of the T. 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 and fight against animal trypanosomosis.