Detection of m(6)A in single cultured cells using scDART-seq

Most techniques for mapping m(6)A-methylated RNAs transcriptome-wide require large amounts of RNA and have been limited to bulk cells and tissues. Here, we provide a detailed protocol for the identification of m(6)A sites in single HEK293T cells using single-cell DART-seq (scDART-seq). The protocol...

Descripción completa

Detalles Bibliográficos
Autores principales: Tegowski, Matthew, Meyer, Kate D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9420395/
https://www.ncbi.nlm.nih.gov/pubmed/36042888
http://dx.doi.org/10.1016/j.xpro.2022.101646
_version_ 1784777379620585472
author Tegowski, Matthew
Meyer, Kate D.
author_facet Tegowski, Matthew
Meyer, Kate D.
author_sort Tegowski, Matthew
collection PubMed
description Most techniques for mapping m(6)A-methylated RNAs transcriptome-wide require large amounts of RNA and have been limited to bulk cells and tissues. Here, we provide a detailed protocol for the identification of m(6)A sites in single HEK293T cells using single-cell DART-seq (scDART-seq). The protocol details how to generate cell lines with inducible expression of the APOBEC1-YTH transgene and the use of important controls for minimizing false positives. We also describe the bioinformatic analysis to identify m(6)A sites. For complete details on the use and execution of this protocol, please refer to Tegowski et al. (2022).
format Online
Article
Text
id pubmed-9420395
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-94203952022-08-29 Detection of m(6)A in single cultured cells using scDART-seq Tegowski, Matthew Meyer, Kate D. STAR Protoc Protocol Most techniques for mapping m(6)A-methylated RNAs transcriptome-wide require large amounts of RNA and have been limited to bulk cells and tissues. Here, we provide a detailed protocol for the identification of m(6)A sites in single HEK293T cells using single-cell DART-seq (scDART-seq). The protocol details how to generate cell lines with inducible expression of the APOBEC1-YTH transgene and the use of important controls for minimizing false positives. We also describe the bioinformatic analysis to identify m(6)A sites. For complete details on the use and execution of this protocol, please refer to Tegowski et al. (2022). Elsevier 2022-08-19 /pmc/articles/PMC9420395/ /pubmed/36042888 http://dx.doi.org/10.1016/j.xpro.2022.101646 Text en © 2022 The Author(s) 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
Tegowski, Matthew
Meyer, Kate D.
Detection of m(6)A in single cultured cells using scDART-seq
title Detection of m(6)A in single cultured cells using scDART-seq
title_full Detection of m(6)A in single cultured cells using scDART-seq
title_fullStr Detection of m(6)A in single cultured cells using scDART-seq
title_full_unstemmed Detection of m(6)A in single cultured cells using scDART-seq
title_short Detection of m(6)A in single cultured cells using scDART-seq
title_sort detection of m(6)a in single cultured cells using scdart-seq
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9420395/
https://www.ncbi.nlm.nih.gov/pubmed/36042888
http://dx.doi.org/10.1016/j.xpro.2022.101646
work_keys_str_mv AT tegowskimatthew detectionofm6ainsingleculturedcellsusingscdartseq
AT meyerkated detectionofm6ainsingleculturedcellsusingscdartseq

Ejemplares similares