ASENCIO, Corinne; HERVÉ, Perrine; MORAND, Pauline; OLIVERES, Quentin; MOREL, Chloé Alexandra; PROUZET-MAULEON, Valérie; BIRAN, Marc; MONIC, Sarah; BONHIVERS, Mélanie; ROBINSON, Derrick Roy; OUELLETTE, Marc; RIVIÈRE, Loïc; BRINGAUD, Frédéric; TETAUD, Emmanuel

doi:10.1111/mmi.15256

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ASENCIO, Corinne
Microbiologie Fondamentale et Pathogénicité [MFP]

HERVÉ, Perrine
Microbiologie Fondamentale et Pathogénicité [MFP]

MORAND, Pauline
Microbiologie Fondamentale et Pathogénicité [MFP]

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Article de revue

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Molecular Microbiology. 2024-06-01, vol. 121, n° 6, p. 1079-1094

Résumé en anglais

Kinetoplastids are unicellular eukaryotic flagellated parasites found in a wide range of hosts within the animal and plant kingdoms. They are known to be responsible in humans for African sleeping sickness (Trypanosoma brucei), Chagas disease (Trypanosoma cruzi), and various forms of leishmaniasis (Leishmania spp.), as well as several animal diseases with important economic impact (African trypanosomes, including Trypanosoma congolense). Understanding the biology of these parasites necessarily implies the ability to manipulate their genomes. In this study, we demonstrate that transfection of a ribonucleoprotein complex, composed of recombinant Streptococcus pyogenes Cas9 (SpCas9) and an in vitro-synthesized guide RNA, results in rapid and efficient genetic modifications of trypanosomatids, in marker-free conditions. This approach was successfully developed to inactivate, delete, and mutate candidate genes in various stages of the life cycle of T. brucei and T. congolense, and Leishmania promastigotes. The functionality of SpCas9 in these parasites now provides, to the research community working on these parasites, a rapid and efficient method of genome editing, without requiring plasmid construction and selection by antibiotics but requires only cloning and PCR screening of the clones. Importantly, this approach is adaptable to any wild-type parasite.< Réduire

Mots clés en anglais

Gene Editing

CRISPR-Cas Systems

Ribonucleoproteins

Streptococcus pyogenes

Leishmania

CRISPR-Associated Protein 9

RNA

Guide

CRISPR-Cas Systems

Trypanosoma brucei brucei

Trypanosoma

Transfection

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https://oskar-bordeaux.fr/handle/20.500.12278/202081

DOI

http://dx.doi.org/10.1111/mmi.15256

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Streptococcus pyogenes Cas9 ribonucleoprotein delivery for efficient, rapid and marker-free gene editing in Trypanosoma and Leishmania.

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Streptococcus pyogenes Cas9 ribonucleoprotein delivery for efficient, rapid and marker-free gene editing in Trypanosoma and Leishmania.

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