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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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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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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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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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RECODE: a programmable guide-free C-to-U RNA editing tool.

Mizuho Ichinose1, Masaru Ohta1, Yasuka Shimajiri1

  • 1EditForce, Inc., Fukuoka 819-0395, Japan.

Nucleic Acids Research
|December 8, 2025
PubMed
Summary
This summary is machine-generated.

A new guide-free RNA editing tool, RECODE, efficiently converts cytidine-to-uridine (C-to-U) in human cells and mice. This programmable RNA editing technology shows therapeutic potential for various diseases.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • CRISPR-based RNA cytidine deaminase tools offer programmable C-to-U editing but face limitations like low efficiency and off-target effects.
  • Existing RNA editing systems require guide RNAs, limiting targetable sequences and flexibility.

Purpose of the Study:

  • To develop a novel, guide-free RNA editing tool for efficient and specific C-to-U conversion.
  • To engineer and optimize a tool for enhanced editing activity, reduced off-target effects, and broad applicability.

Main Methods:

  • Engineered RECODE (RNA Editor for C-to-U with an Optimized DYW Enzyme) using RNA-binding pentatricopeptide repeat proteins fused to a DYW cytidine deaminase domain.
  • Optimized RECODE's specificity domain for retargeting and reduced off-target edits, and enhanced its catalytic region for increased activity and translation.

Main Results:

  • RECODE demonstrated efficient C-to-U editing in human cells across diverse targets, achieving over 50% efficiency for most sites.
  • The tool showed high specificity, avoiding edits at adjacent cytidines, and proved functional in mice with high efficiency in skeletal muscle tissue via AAV delivery.

Conclusions:

  • RECODE represents a significant advancement in RNA editing technology, offering a guide-free, efficient, and specific C-to-U conversion system.
  • The demonstrated *in vivo* functionality and therapeutic potential suggest RECODE could be valuable for treating various diseases.