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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Site-Specific m6 A Erasing via Conditionally Stabilized CRISPR-Cas13b Editor.

Ying Xu1, Yufan Wang1, Fu-Sen Liang1

  • 1Department of Chemistry, Case Western Reserve University, 2080 Adelbert Rd, Cleveland, OH 44106, USA.

Angewandte Chemie (International Ed. in English)
|September 15, 2023
PubMed
Summary

Researchers developed a conditional RNA editing platform using a ligand-stabilized dCas13 editor for precise N6-methyladenosine (m6 A) modification. This system allows inducible control over m6 A erasure, minimizing steric interference for functional studies.

Keywords:
Cas13bProtein DegradationRNAShield-1m6A

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Area of Science:

  • Molecular Biology
  • RNA Epigenetics
  • Gene Editing Technologies

Background:

  • N6-methyladenosine (m6 A) is a crucial RNA modification regulating diverse biological processes.
  • CRISPR technology enables programmable m6 A editing, but large protein size and constitutive expression pose challenges.
  • Existing CRISPR/RNA editing enzymes can interfere with native RNA and cellular functions.

Purpose of the Study:

  • To develop a conditional and ligand-inducible platform for programmable m6 A editing.
  • To overcome the limitations of bulky CRISPR proteins and constitutive enzyme expression in RNA editing.
  • To enable site-specific m6 A erasure with minimal steric interference for functional studies.

Main Methods:

  • Development of a conditional m6 A editing platform (FKBP*-dCas13b-ALK) utilizing a ligand-stabilized dCas13 editor.
  • Inducible expression of the m6 A editing system controlled by the addition or removal of the Shield-1 molecule.
  • Demonstration of targeted recruitment of dCas13b-m6 A eraser fusion protein and site-specific m6 A erasing under Shield-1 control.

Main Results:

  • Successful implementation of a conditional, ligand-inducible m6 A editing system.
  • Shield-1 molecule effectively controlled the expression and recruitment of the dCas13b fusion protein for targeted m6 A erasure.
  • Rapid release and degradation of the dCas13b fusion protein post-Shield-1 removal, preceding m6 A restoration.

Conclusions:

  • The FKBP*-dCas13b-ALK platform provides a novel method for conditional and inducible m6 A editing.
  • This system allows precise control over m6 A erasure, facilitating the study of m6 A functions.
  • The rapid clearance of the fusion protein minimizes steric hindrance, offering an improved approach for investigating RNA modification roles.