Literature DB >> 36042888

Detection of m6A in single cultured cells using scDART-seq.

Matthew Tegowski1, Kate D Meyer2.   

Abstract

Most techniques for mapping m6A-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 m6A 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 m6A sites. For complete details on the use and execution of this protocol, please refer to Tegowski et al. (2022).
© 2022 The Author(s).

Entities:  

Keywords:  Cell-based Assays; Flow Cytometry/Mass Cytometry; Gene Expression; Molecular Biology; RNAseq; Sequencing; Single Cell

Mesh:

Substances:

Year:  2022        PMID: 36042888      PMCID: PMC9420395          DOI: 10.1016/j.xpro.2022.101646

Source DB:  PubMed          Journal:  STAR Protoc        ISSN: 2666-1667


  9 in total

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Authors:  Yang Liao; Gordon K Smyth; Wei Shi
Journal:  Bioinformatics       Date:  2013-11-13       Impact factor: 6.937

2.  Comprehensive Integration of Single-Cell Data.

Authors:  Tim Stuart; Andrew Butler; Paul Hoffman; Christoph Hafemeister; Efthymia Papalexi; William M Mauck; Yuhan Hao; Marlon Stoeckius; Peter Smibert; Rahul Satija
Journal:  Cell       Date:  2019-06-06       Impact factor: 41.582

3.  STAR: ultrafast universal RNA-seq aligner.

Authors:  Alexander Dobin; Carrie A Davis; Felix Schlesinger; Jorg Drenkow; Chris Zaleski; Sonali Jha; Philippe Batut; Mark Chaisson; Thomas R Gingeras
Journal:  Bioinformatics       Date:  2012-10-25       Impact factor: 6.937

4.  scDART-seq reveals distinct m6A signatures and mRNA methylation heterogeneity in single cells.

Authors:  Matthew Tegowski; Mathieu N Flamand; Kate D Meyer
Journal:  Mol Cell       Date:  2022-01-25       Impact factor: 17.970

5.  The Sequence Alignment/Map format and SAMtools.

Authors:  Heng Li; Bob Handsaker; Alec Wysoker; Tim Fennell; Jue Ruan; Nils Homer; Gabor Marth; Goncalo Abecasis; Richard Durbin
Journal:  Bioinformatics       Date:  2009-06-08       Impact factor: 6.937

6.  BEDTools: a flexible suite of utilities for comparing genomic features.

Authors:  Aaron R Quinlan; Ira M Hall
Journal:  Bioinformatics       Date:  2010-01-28       Impact factor: 6.937

7.  Flexbar 3.0 - SIMD and multicore parallelization.

Authors:  Johannes T Roehr; Christoph Dieterich; Knut Reinert
Journal:  Bioinformatics       Date:  2017-09-15       Impact factor: 6.937

8.  MetaPlotR: a Perl/R pipeline for plotting metagenes of nucleotide modifications and other transcriptomic sites.

Authors:  Anthony O Olarerin-George; Samie R Jaffrey
Journal:  Bioinformatics       Date:  2017-05-15       Impact factor: 6.937

9.  DART-seq: an antibody-free method for global m6A detection.

Authors:  Kate D Meyer
Journal:  Nat Methods       Date:  2019-09-23       Impact factor: 28.547

  9 in total

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