Cargando…
High-Resolution Single-Cell Sequencing of Malaria Parasites
Single-cell genomics is a powerful tool for determining the genetic architecture of complex communities of unicellular organisms. In areas of high transmission, malaria patients are often challenged by the activities of multiple Plasmodium falciparum lineages, which can potentiate pathology, spread...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5737330/ https://www.ncbi.nlm.nih.gov/pubmed/29220419 http://dx.doi.org/10.1093/gbe/evx256 |
_version_ | 1783287501055590400 |
---|---|
author | Trevino, Simon G Nkhoma, Standwell C Nair, Shalini Daniel, Benjamin J Moncada, Karla Khoswe, Stanley Banda, Rachel L Nosten, François Cheeseman, Ian H |
author_facet | Trevino, Simon G Nkhoma, Standwell C Nair, Shalini Daniel, Benjamin J Moncada, Karla Khoswe, Stanley Banda, Rachel L Nosten, François Cheeseman, Ian H |
author_sort | Trevino, Simon G |
collection | PubMed |
description | Single-cell genomics is a powerful tool for determining the genetic architecture of complex communities of unicellular organisms. In areas of high transmission, malaria patients are often challenged by the activities of multiple Plasmodium falciparum lineages, which can potentiate pathology, spread drug resistance loci, and also complicate most genetic analysis. Single-cell sequencing of P. falciparum would be key to understanding infection complexity, though efforts are hampered by the extreme nucleotide composition of its genome (∼80% AT-rich). To counter the low coverage achieved in previous studies, we targeted DNA-rich late-stage parasites by Fluorescence-Activated Cell Sorting and whole genome sequencing. Our method routinely generates accurate, near-complete capture of the 23 Mb P. falciparum genome (mean breadth of coverage 90.7%) at high efficiency. Data from 48 single-cell genomes derived from a polyclonal infection sampled in Chikhwawa, Malawi allowed for unambiguous determination of haplotype diversity and recent meiotic events, information that will aid public health efforts. |
format | Online Article Text |
id | pubmed-5737330 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-57373302018-01-08 High-Resolution Single-Cell Sequencing of Malaria Parasites Trevino, Simon G Nkhoma, Standwell C Nair, Shalini Daniel, Benjamin J Moncada, Karla Khoswe, Stanley Banda, Rachel L Nosten, François Cheeseman, Ian H Genome Biol Evol Research Article Single-cell genomics is a powerful tool for determining the genetic architecture of complex communities of unicellular organisms. In areas of high transmission, malaria patients are often challenged by the activities of multiple Plasmodium falciparum lineages, which can potentiate pathology, spread drug resistance loci, and also complicate most genetic analysis. Single-cell sequencing of P. falciparum would be key to understanding infection complexity, though efforts are hampered by the extreme nucleotide composition of its genome (∼80% AT-rich). To counter the low coverage achieved in previous studies, we targeted DNA-rich late-stage parasites by Fluorescence-Activated Cell Sorting and whole genome sequencing. Our method routinely generates accurate, near-complete capture of the 23 Mb P. falciparum genome (mean breadth of coverage 90.7%) at high efficiency. Data from 48 single-cell genomes derived from a polyclonal infection sampled in Chikhwawa, Malawi allowed for unambiguous determination of haplotype diversity and recent meiotic events, information that will aid public health efforts. Oxford University Press 2017-12-06 /pmc/articles/PMC5737330/ /pubmed/29220419 http://dx.doi.org/10.1093/gbe/evx256 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Trevino, Simon G Nkhoma, Standwell C Nair, Shalini Daniel, Benjamin J Moncada, Karla Khoswe, Stanley Banda, Rachel L Nosten, François Cheeseman, Ian H High-Resolution Single-Cell Sequencing of Malaria Parasites |
title | High-Resolution Single-Cell Sequencing of Malaria Parasites |
title_full | High-Resolution Single-Cell Sequencing of Malaria Parasites |
title_fullStr | High-Resolution Single-Cell Sequencing of Malaria Parasites |
title_full_unstemmed | High-Resolution Single-Cell Sequencing of Malaria Parasites |
title_short | High-Resolution Single-Cell Sequencing of Malaria Parasites |
title_sort | high-resolution single-cell sequencing of malaria parasites |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5737330/ https://www.ncbi.nlm.nih.gov/pubmed/29220419 http://dx.doi.org/10.1093/gbe/evx256 |
work_keys_str_mv | AT trevinosimong highresolutionsinglecellsequencingofmalariaparasites AT nkhomastandwellc highresolutionsinglecellsequencingofmalariaparasites AT nairshalini highresolutionsinglecellsequencingofmalariaparasites AT danielbenjaminj highresolutionsinglecellsequencingofmalariaparasites AT moncadakarla highresolutionsinglecellsequencingofmalariaparasites AT khoswestanley highresolutionsinglecellsequencingofmalariaparasites AT bandarachell highresolutionsinglecellsequencingofmalariaparasites AT nostenfrancois highresolutionsinglecellsequencingofmalariaparasites AT cheesemanianh highresolutionsinglecellsequencingofmalariaparasites |