Cargando…

Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations

Adaptive T and B lymphocytes expand, respond, and persist across a multitude of separable cell differentiation states. Small compartments of these cells present defined cell surface phenotype, but express potentially divergent immune functions. Here, we use high resolution flow cytometry to provide...

Descripción completa

Detalles Bibliográficos
Autores principales: Shuparski, Andrew G., Higgins, Brett W., Miller, Karen B., Dufaud, Chad R., Robinson, Amanda M., Dueker, Katharina, McHeyzer-Williams, Louise J., McHeyzer-Williams, Michael G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715333/
https://www.ncbi.nlm.nih.gov/pubmed/35005646
http://dx.doi.org/10.1016/j.xpro.2021.101064
_version_ 1784624108243255296
author Shuparski, Andrew G.
Higgins, Brett W.
Miller, Karen B.
Dufaud, Chad R.
Robinson, Amanda M.
Dueker, Katharina
McHeyzer-Williams, Louise J.
McHeyzer-Williams, Michael G.
author_facet Shuparski, Andrew G.
Higgins, Brett W.
Miller, Karen B.
Dufaud, Chad R.
Robinson, Amanda M.
Dueker, Katharina
McHeyzer-Williams, Louise J.
McHeyzer-Williams, Michael G.
author_sort Shuparski, Andrew G.
collection PubMed
description Adaptive T and B lymphocytes expand, respond, and persist across a multitude of separable cell differentiation states. Small compartments of these cells present defined cell surface phenotype, but express potentially divergent immune functions. Here, we use high resolution flow cytometry to provide direct access to rare lymphocyte subpopulations for evaluation of steady-state or reactive transcriptional programs. We sort and index single cells by phenotype in 384-well format for quantification of targeted gene amplification through RNA sequencing (single cell qtSEQ). For complete details on the use and execution of this profile, please refer to Dufaud et al. (2021).
format Online
Article
Text
id pubmed-8715333
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-87153332022-01-06 Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations Shuparski, Andrew G. Higgins, Brett W. Miller, Karen B. Dufaud, Chad R. Robinson, Amanda M. Dueker, Katharina McHeyzer-Williams, Louise J. McHeyzer-Williams, Michael G. STAR Protoc Protocol Adaptive T and B lymphocytes expand, respond, and persist across a multitude of separable cell differentiation states. Small compartments of these cells present defined cell surface phenotype, but express potentially divergent immune functions. Here, we use high resolution flow cytometry to provide direct access to rare lymphocyte subpopulations for evaluation of steady-state or reactive transcriptional programs. We sort and index single cells by phenotype in 384-well format for quantification of targeted gene amplification through RNA sequencing (single cell qtSEQ). For complete details on the use and execution of this profile, please refer to Dufaud et al. (2021). Elsevier 2021-12-22 /pmc/articles/PMC8715333/ /pubmed/35005646 http://dx.doi.org/10.1016/j.xpro.2021.101064 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Shuparski, Andrew G.
Higgins, Brett W.
Miller, Karen B.
Dufaud, Chad R.
Robinson, Amanda M.
Dueker, Katharina
McHeyzer-Williams, Louise J.
McHeyzer-Williams, Michael G.
Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title_full Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title_fullStr Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title_full_unstemmed Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title_short Single cell qtSEQ: Cell-indexed quantitative and targeted RNA sequencing for sorted rare lymphocyte subpopulations
title_sort single cell qtseq: cell-indexed quantitative and targeted rna sequencing for sorted rare lymphocyte subpopulations
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715333/
https://www.ncbi.nlm.nih.gov/pubmed/35005646
http://dx.doi.org/10.1016/j.xpro.2021.101064
work_keys_str_mv AT shuparskiandrewg singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT higginsbrettw singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT millerkarenb singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT dufaudchadr singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT robinsonamandam singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT duekerkatharina singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT mcheyzerwilliamslouisej singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations
AT mcheyzerwilliamsmichaelg singlecellqtseqcellindexedquantitativeandtargetedrnasequencingforsortedrarelymphocytesubpopulations