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Related Concept Videos

RNA Interference01:23

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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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RNA Editing02:23

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Related Experiment Video

Updated: Sep 6, 2025

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins
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Sequence-specific targeting of RNA.

Lukas Rösner1, Dennis Reichert2, Kristina Rau1

  • 1University of Münster, Department of Chemistry, Institute of Biochemistry, Corrensstraße 36, 48149 Münster, Germany.

Methods (San Diego, Calif.)
|June 28, 2022
PubMed
Summary

Researchers developed novel tools to precisely add or remove RNA methylations, crucial for gene expression regulation. This advancement enables specific targeting of RNA modifications like N7-methylation and N6-methyladenosine (m6A) in eukaryotic cells.

Keywords:
Ecm1FTOMethyltransferaseN(6)-methyladenosine (m(6)A)RNA modificationRNA-binding proteins

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • RNA Biology

Background:

  • Post-transcriptional modifications regulate vital cellular processes like translation and RNA degradation.
  • Understanding specific RNA modifications requires tools for sequence-specific RNA targeting.

Purpose of the Study:

  • To develop and demonstrate tools for precise, sequence-specific installation and removal of key mRNA modifications.
  • To investigate the roles of N7-methylation of the 5'-cap and N6-methyladenosine (m6A) modifications.

Main Methods:

  • Utilized nuclease-deficient RNA-targeting Cas9 (RCas9) for sequence-specific RNA targeting.
  • Employed Pumilio homology domain (PumHD) fused to effector enzymes (FTO, Ecm1) for targeted modification.
  • Applied these tools to model RNA transcripts to install and remove methylations.

Main Results:

  • Successfully achieved sequence-specific targeting of RNA transcripts.
  • Demonstrated the ability to install and remove N7-methylation and m6A modifications with high specificity.
  • Validated the efficacy of RCas9 and PumHD-based systems for RNA modification control.

Conclusions:

  • Developed a versatile toolkit for precise manipulation of RNA methylation.
  • These tools facilitate functional studies of specific RNA modifications in eukaryotic systems.
  • Opens new avenues for understanding gene regulation at the post-transcriptional level.