Bioluminescent-fluorescent T. cruzi for Chagas Disease Research

Trypanosoma cruzi is the causative agent of Chagas disease, a major public health problem in Latin America and an emergent disease in the US. There is no vaccine available, and drugs are limited with severe side effects. Efforts to generate new therapies are affected by limitations in the understanding of parasite biology and disease pathogenesis [1]. Until recently, the functional dissection of T. cruzi biology has been restricted by limited tools of genetic manipulation of the parasite [2]. BEI Resources now offers a transgenic reporter strain of T. cruzi (NR-59633) that expresses a fusion protein comprised of a red-shifted variant of the firefly luciferase gene (PpyRE9h) and green fluorescent protein (mNeonGreen). Bioluminescence enables real-time tracking of infection dynamics in vivo, while the fluorescent marker allows for visualization of individual parasites in tissue sections, facilitating studies of host-parasite interactions at the cellular level [3, 4]. Additionally, this strain incorporates a streamlined CRISPR/Cas9 system that supports genome editing through a PCR-based method, eliminating the need for DNA cloning. The system provided by this genetically engineered T. cruzi strain allows the rapid generation of null mutants and fluorescently tagged parasites in a background where the in vivo phenotype can be rapidly assessed.

References:

1. de Sousa A. S. et al. “Chagas Disease.” Lancet. 403 (2024):203-218. PubMed: 38071985.

2. de Assis Burle-Caldas, G., et al. “CRISPR Genome Editing and the Study of Chagas Disease.” Adv. Exp. Med. Biol. 1429 (2023): 111-125. PubMed: 37486519. .

3. Costa, F. C., et al. Expanding the Toolbox for Trypanosoma Cruzi: A Parasite Line Incorporating a Bioluminescence-Fluorescence Dual Reporter and Streamlined CRISPR/Cas9 Functionality for Rapid In Vivo Localisation and Phenotyping. PLoS Negl. Trop. Dis. 12 (2018): e0006388. PubMed: 29608569.

4. Olmo, F., et al. “A Panel of Phenotypically and Genotypically Diverse Bioluminescent:Fluorescent Trypanosoma Cruzi Strains as a Resource for Chagas Disease Research.” PLoS Negl. Trop. Dis. 18 (2024): e0012106. PubMed: 38820564.

Image Alt Text: T. Cruzi in brightfield and mNeonGreen Credit: BEI Resources

Image Credit: T. Cruzi in brightfield and mNeonGreen Credit: BEI Resources