High‑throughput analysis of the Trypanosoma cruzi minicirculome (mcDNA) unveils structural variation and functional diversity

Abstract

Trypanosoma cruzi causes Chagas disease and has a unique extranuclear genome enclosed in a structure called the kinetoplast, which contains circular genomes known as maxi‑ and minicircles. While the structure and function of maxicircles are well‑understood, many aspects of minicircles remain to be discovered. Here, we performed a high‑throughput analysis of the minicirculome (mcDNA) in 50 clones isolated from Colombia’s diverse T. cruzi I populations. Results indicate that mcDNA comprises four diverse subpopulations with different structures, lengths, and numbers of interspersed semi‑conserved (previously termed ultra‑conserved regions mHCV) and hypervariable (mHVPs) regions. Analysis of mcDNA ancestry and inter‑clone differentiation indicates the interbreeding of minicircle sequence classes is placed along diverse strains and hosts. These results support evidence of the multiclonal dynamics and random bi‑parental segregation. Finally, we disclosed the guide RNA repertoire encoded by mcDNA at a clonal scale, and several attributes of its abundance and function are discussed.