Cargando…
MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling
Recently developed single-cell RNA sequencing methods allow the simultaneous profiling of the transcriptomes of thousands of individual cells. However, current methods still require advanced equipment or entail substantial waste of reagents. Here, we introduce magnetic bead-assisted parallel single-...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7272464/ https://www.ncbi.nlm.nih.gov/pubmed/32404928 http://dx.doi.org/10.1038/s12276-020-0433-x |
_version_ | 1783542260363689984 |
---|---|
author | Park, Munsu Lee, Dongin Bang, Duhee Lee, Ji Hyun |
author_facet | Park, Munsu Lee, Dongin Bang, Duhee Lee, Ji Hyun |
author_sort | Park, Munsu |
collection | PubMed |
description | Recently developed single-cell RNA sequencing methods allow the simultaneous profiling of the transcriptomes of thousands of individual cells. However, current methods still require advanced equipment or entail substantial waste of reagents. Here, we introduce magnetic bead-assisted parallel single-cell gene expression sequencing (MAPS-seq), a microwell-based method that pools samples before the reverse transcription step, increasing the ease of sample preparation and reducing reagent waste. Moreover, because this method uses universal reagents and standard molecular biology lab instruments, it is easy to implement, even in labs that have not previously conducted single-cell RNA sequencing. We validated our method by demonstrating that it can generate gene expression data at the single-cell level. We then applied the MAPS-seq method to analyze 237 human myelogenous leukemia cells treated with one of three different drugs or dimethyl sulfoxide. We observed transcriptional changes and identified marker genes that indicate a drug response. Furthermore, the MAPS-seq method produced data of comparable quality to those of existing single-cell RNA sequencing methods. Consequently, we expect that our method will provide researchers with a more accessible, less wasteful, and less burdensome method for investigating the transcriptomes of individual cells. |
format | Online Article Text |
id | pubmed-7272464 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-72724642020-06-15 MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling Park, Munsu Lee, Dongin Bang, Duhee Lee, Ji Hyun Exp Mol Med Article Recently developed single-cell RNA sequencing methods allow the simultaneous profiling of the transcriptomes of thousands of individual cells. However, current methods still require advanced equipment or entail substantial waste of reagents. Here, we introduce magnetic bead-assisted parallel single-cell gene expression sequencing (MAPS-seq), a microwell-based method that pools samples before the reverse transcription step, increasing the ease of sample preparation and reducing reagent waste. Moreover, because this method uses universal reagents and standard molecular biology lab instruments, it is easy to implement, even in labs that have not previously conducted single-cell RNA sequencing. We validated our method by demonstrating that it can generate gene expression data at the single-cell level. We then applied the MAPS-seq method to analyze 237 human myelogenous leukemia cells treated with one of three different drugs or dimethyl sulfoxide. We observed transcriptional changes and identified marker genes that indicate a drug response. Furthermore, the MAPS-seq method produced data of comparable quality to those of existing single-cell RNA sequencing methods. Consequently, we expect that our method will provide researchers with a more accessible, less wasteful, and less burdensome method for investigating the transcriptomes of individual cells. Nature Publishing Group UK 2020-05-13 /pmc/articles/PMC7272464/ /pubmed/32404928 http://dx.doi.org/10.1038/s12276-020-0433-x Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Park, Munsu Lee, Dongin Bang, Duhee Lee, Ji Hyun MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title | MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title_full | MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title_fullStr | MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title_full_unstemmed | MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title_short | MAPS-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
title_sort | maps-seq: magnetic bead-assisted parallel single-cell gene expression profiling |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7272464/ https://www.ncbi.nlm.nih.gov/pubmed/32404928 http://dx.doi.org/10.1038/s12276-020-0433-x |
work_keys_str_mv | AT parkmunsu mapsseqmagneticbeadassistedparallelsinglecellgeneexpressionprofiling AT leedongin mapsseqmagneticbeadassistedparallelsinglecellgeneexpressionprofiling AT bangduhee mapsseqmagneticbeadassistedparallelsinglecellgeneexpressionprofiling AT leejihyun mapsseqmagneticbeadassistedparallelsinglecellgeneexpressionprofiling |