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Targeted m6A reader proteins to study the epitranscriptome.

Simone Rauch1, Bryan C Dickinson2

  • 1Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, United States; Department of Chemistry, The University of Chicago, Chicago, IL, United States.

Methods in Enzymology
|May 27, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed programmable reader proteins to study RNA modifications like N-methyladenosine (m6A) at the single transcript level. This advances understanding of epitranscriptomic regulation in eukaryotic systems.

Keywords:
Cas13N(6)-methyltranscriptomeProtein engineeringRNA modificationsSynthetic biology

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Posttranscriptional regulation of RNA is crucial for genetic information flow in eukaryotes.
  • Chemical modifications on RNA, particularly N6-methyladenosine (m6A) in mammals, impact mRNA lifetime, location, trafficking, and function.
  • While m6A regulatory proteins (writers, erasers, readers) are known at the cellular level, single-transcript analysis is needed.

Purpose of the Study:

  • To detail the application of targeted m6A reader proteins for studying epitranscriptomic regulation at single endogenous RNA sites.
  • To provide a protocol for investigating RNA modifications using programmable reader proteins.

Main Methods:

  • Development of programmable m6A reader proteins.
  • Utilizing Cas13 RNA-targeting proteins.
  • Application of targeted readers to study RNA regulation at single endogenous sites.

Main Results:

  • Demonstrated the utility of programmable m6A readers for single-transcript epitranscriptomic analysis.
  • Established a protocol for studying RNA modifications with targeted reader proteins.

Conclusions:

  • Targeted m6A readers offer a powerful tool to investigate epitranscriptomic regulation at the single transcript level.
  • This approach is key to understanding the mechanisms and consequences of m6A modification in gene regulation.