Title
Evaluation of whole genome amplification and bioinformatic methods for the characterization of Leishmania genomes at a single cell level
Date Issued
01 December 2020
Access level
open access
Resource Type
journal article
Author(s)
Imamura H.
Monsieurs P.
Sanders M.
Maes I.
Vanaerschot M.
Berriman M.
Cotton J.A.
Dujardin J.C.
Domagalska M.A.
Institute of Tropical Medicine Antwerp
Publisher(s)
Nature Research
Abstract
Here, we report a pilot study paving the way for further single cell genomics studies in Leishmania. First, the performances of two commercially available kits for Whole Genome Amplification (WGA), PicoPLEX and RepliG were compared on small amounts of Leishmania donovani DNA, testing their ability to preserve specific genetic variations, including aneuploidy levels and SNPs. We show here that the choice of WGA method should be determined by the planned downstream genetic analysis, PicoPLEX and RepliG performing better for aneuploidy and SNP calling, respectively. This comparison allowed us to evaluate and optimize corresponding bio-informatic methods. As PicoPLEX was shown to be the preferred method for studying single cell aneuploidy, this method was applied in a second step, on single cells of L. braziliensis, which were sorted by fluorescence activated cell sorting (FACS). Even sequencing depth was achieved in 28 single cells, allowing accurate somy estimation. A dominant karyotype with three aneuploid chromosomes was observed in 25 cells, while two different minor karyotypes were observed in the other cells. Our method thus allowed the detection of aneuploidy mosaicism, and provides a solid basis which can be further refined to concur with higher-throughput single cell genomic methods.
Volume
10
Issue
1
Language
English
OCDE Knowledge area
BioinformĂ¡tica
IngenierĂa elĂ©ctrica, IngenierĂa electrĂ³nica
Scopus EID
2-s2.0-85090945478
PubMed ID
Source
Scientific Reports
ISSN of the container
20452322
Sponsor(s)
This study received financial support from the Flemish Ministry of Science and Innovation (SOFI Grants SINGLE and MADLEI), the Flemish Fund for Scientific Research (FWO, post-doctoral grant to MJ). JAC, MB and MS were supported by Wellcome via the core support of the Wellcome Sanger Institute (grant 206194).
Sources of information:
Directorio de ProducciĂ³n CientĂfica
Scopus