Identification and analysis of the DNA content of small extracellular vesicles isolated from Leishmania parasites

Ana Victoria Ibarra-Meneses; Atia Amin; George Dong; Martin Olivier; David Langlais; Christopher Fernandez-Prada

doi:10.1016/j.xpro.2023.102248

Identification and analysis of the DNA content of small extracellular vesicles isolated from Leishmania parasites

STAR Protoc. 2023 Apr 21;4(2):102248. doi: 10.1016/j.xpro.2023.102248. Online ahead of print.

Authors

Ana Victoria Ibarra-Meneses¹, Atia Amin², George Dong³, Martin Olivier⁴, David Langlais⁵, Christopher Fernandez-Prada⁶

Affiliations

¹ Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Université de Montréal, 626 CIMIA Sicotte Street, Saint-Hyacinthe, QC J2S 2M2, Canada; The Research Group on Infectious Diseases in Production Animals (GREMIP), Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC J2S 2M2, Canada.
² Department of Human Genetics, Dahdaleh Institute for Genomic Medicine, Montreal, QC H3A 0G1, Canada.
³ IDIGH, The Research Institute of the McGill University Health Centre, 2155 Guy Street, Montreal, QC H3H 2L9, Canada.
⁴ IDIGH, The Research Institute of the McGill University Health Centre, 2155 Guy Street, Montreal, QC H3H 2L9, Canada; Department of Microbiology and Immunology, McGill Research Centre on Complex Traits, Montreal, QC, Canada. Electronic address: martin.olivier@mcgill.ca.
⁵ Department of Human Genetics, Dahdaleh Institute for Genomic Medicine, Montreal, QC H3A 0G1, Canada; Department of Microbiology and Immunology, McGill Research Centre on Complex Traits, Montreal, QC, Canada. Electronic address: david.langlais@mcgill.ca.
⁶ Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Université de Montréal, 626 CIMIA Sicotte Street, Saint-Hyacinthe, QC J2S 2M2, Canada; The Research Group on Infectious Diseases in Production Animals (GREMIP), Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC J2S 2M2, Canada; Department of Microbiology and Immunology, McGill Research Centre on Complex Traits, Montreal, QC, Canada. Electronic address: christopher.fernandez.prada@umontreal.ca.

Abstract

Here, we focus on Leishmania extracellular vesicles (EVs) and their DNA content, detailing a protocol for the isolation of these nanoparticles and their subsequent genomic characterization. We describe a robust and comprehensive approach for obtaining, storing, and analyzing EVs derived from cultured parasites. We detail a user-friendly bioinformatics pipeline for sequence analysis and visualization of CNV analysis and ploidy changes. For complete details on the use and execution of this protocol, please refer to Douanne et al. (2022).¹.

Keywords: Cell Biology; Genomics; Microbiology; Microscopy; Sequence Analysis; Sequencing.