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...

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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
Descripción
Sumario: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).

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