Gabriel Negreira

A novel high-throughput single-cell DNA sequencing method reveals hidden genomic heterogeneity in the unicellular eukaryote Leishmania Genome instability is considered a major driver of adaptation in eukaryotic microorganisms, but its study is hampered by the limited availability of genomic technologies with single-cell resolution. H...

This is just the beginning! We hope this method will help the community to explore genome evolution and parasite adaptation in Leishmania and beyond!

For more info, check the preprint in @biorxivpreprint.bsky.social:

www.biorxiv.org/content/10.1...

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We also revealed a previously unknown population heterogeneity in a #Leishmania strain which is commonly used in laboratory research.

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In our latest preprint, we combined Semi-permeable capsules ( #SPCs ) with #PTA to sequence the DNA of individual #Leishmania parasites at high throughput, identifying multiple layers of genetic heterogeneity, from mosaic aneuploidy to variations in the sequence of drug-resistance genes.

Single Cell CNV | 10x Genomics Chromium Single Cell CNV provides a comprehensive, scalable solution for revealing genome heterogeneity and understanding clonal evolution. Study disease pathogenesis or characterize neuronal mosaicis...

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However, the Chromium single-cell CNV solution was discontinued in 2020, and since then, the filed has been left without viable alternatives.

www.10xgenomics.com/products/sin...

The adaptive roles of aneuploidy and polyclonality in Leishmania in response to environmental stress | EMBO reports image image The dynamics of aneuploidy, a hallmark of Leishmania adaptation, depends on the nature and strength of the environmental stress as well as on the existence of other pre‐adaptive mechanism...

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We also used it to understand how karyotype diversity (mosaic aneuploidy) could help parasites to survive drugs that in principle should kill them:

www.embopress.org/doi/full/10....

High throughput single-cell genome sequencing gives insights into the generation and evolution of mosaic aneuploidy in Leishmania donovani Abstract. Leishmania, a unicellular eukaryotic parasite, is a unique model for aneuploidy and cellular heterogeneity, along with their potential role in ad

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Later, we used the Chromium Single-cell CNV solution (10X Genomic) to determine the chromosome configuration (karyotypes) of thousands of Leishmania parasites, which led to some hypothesis on how karyotypes evolve in culture.

academic.oup.com/nar/article/...

Evaluation of whole genome amplification and bioinformatic methods for the characterization of Leishmania genomes at a single cell level - Scientific Reports Scientific Reports - Evaluation of whole genome amplification and bioinformatic methods for the characterization of Leishmania genomes at a single cell level

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More than 5 years ago we embarked on the journey of using #scDNAseq to study genetic heterogeneity in #Leishmania.

www.nature.com/articles/s41...

Happy to share our latest preprint, where we present a novel high-throughput #single-cell DNA method that resolves genomic heterogeneity in #Leishmania, from #aneuploidy, #CNVs to #drug-resistance mutations.

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Link here: www.biorxiv.org/content/10.1...

Drs Kyrie Dickson, Jaime Costales, Malgorzata Domagalska, Ferre Veken & Martin Llewellyn discuss #genome (dis)organisation in #Trypanosoma #cruzi: innovation through #instability? #Adaptation #Chagas #Aneuploidy #Trypanosomatids

www.cell.com/trends/paras...

R-loops acted on by RNase H1 influence DNA replication timing and genome stability in Leishmania - Nature Communications All genomes are replicated in a temporal order, though the determinants of such timing are poorly characterized. Here, the authors show that the unusual chromosome size-dependent DNA replication timin...

Please enjoy a new paper led by @jezieldamasceno.bsky.social that reveals how R-loops and their homeostatsis through RNase H1 modulate Leishmania DNA replication timing @uofgsii.bsky.social @marquescata.bsky.social @scotomorph.bsky.social
www.nature.com/articles/s41...

🌍Did you know visceral leishmaniasis claims thousands of lives each year in East Africa? In light of this World NTD Day, ITM alumnus Ermias Diro will shed light on the challenges and solutions to eliminate this disease. Reserve your spot today!👉 bit.ly/3PJfzM0 #EndNTDs