cris.boxmetadata.label.title
Evaluation of whole genome amplification and bioinformatic methods for the characterization of Leishmania genomes at a single cell level
cris.boxmetadata.label.dateissued
01 browse.startsWith.months.december 2020
cris.boxmetadata.label.accesslevel
open access
cris.boxmetadata.label.resourcetype
journal article
cris.boxmetadata.label.authors
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
cris.boxmetadata.label.publisher
Nature Research
cris.boxmetadata.label.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.
cris.boxmetadata.label.volume
10
cris.boxmetadata.label.issue
1
cris.boxmetadata.label.language
English
cris.boxmetadata.label.ocdeknowledgeArea
IngenierĂ­a elĂ©ctrica, IngenierĂ­a electrĂ³nica BioinformĂ¡tica
cris.boxmetadata.label.doi
cris.boxmetadata.label.scopusidentifier
2-s2.0-85090945478
cris.boxmetadata.label.pubmedidentifier
cris.boxmetadata.label.source
Scientific Reports
cris.boxmetadata.label.containerissn
20452322
cris.boxmetadata.label.sponsor
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).
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