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Related Concept Videos

CRISPR01:59

CRISPR

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 Short...
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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Updated: Jun 12, 2026

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Enhanced RNA-targeting CRISPR-Cas technology in zebrafish.

Ismael Moreno-Sanchez1,2,3, Luis Hernandez-Huertas1,2, Daniel Nahon-Cano1,2

  • 1Andalusian Center for Developmental Biology (CABD), Pablo de Olavide University/CSIC/Junta de Andalucía, Ctra. Utrera Km.1, 41013, Seville, Spain.

Biorxiv : the Preprint Server for Biology
|October 17, 2024
PubMed
Summary

Optimized CRISPR-RfxCas13d RNA targeting in zebrafish using transient formulations. This enhanced system efficiently depletes mRNA in embryos with minimal collateral effects, advancing in vivo applications.

Keywords:
CRISPR-CasCas13Cas7–11RNA-targetingcollateral activityzebrafish

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas13 systems are valuable tools but face challenges with collateral activity and in vivo efficiency.
  • Optimizing RNA targeting is crucial for advancing CRISPR applications in biological research.

Purpose of the Study:

  • To enhance the CRISPR-RfxCas13d system for efficient and specific RNA targeting in zebrafish.
  • To investigate transient formulations and chemical modifications for improved in vivo performance.
  • To develop predictive models for guide RNA activity and minimize off-target effects.

Main Methods:

  • Utilized chemically modified guide RNAs (gRNAs) for enhanced loss-of-function phenotypes.
  • Improved nuclear RNA targeting strategies within the CRISPR-RfxCas13d system.
  • Compared computational models to predict in vivo gRNA activity accurately.
  • Implemented alternative RNA-targeting CRISPR-Cas systems with reduced collateral activity.

Main Results:

  • Transient CRISPR-RfxCas13d formulations (ribonucleoprotein complexes or mRNA-gRNA) significantly enhanced system efficiency in zebrafish.
  • Chemically modified gRNAs led to more penetrant loss-of-function phenotypes.
  • Accurate prediction of gRNA activity in vivo was achieved using optimized computational models.
  • CRISPR-RfxCas13d effectively depleted endogenous mRNAs in zebrafish embryos with minimal collateral effects, except for extremely abundant or ectopic RNAs.

Conclusions:

  • Transient CRISPR-Cas13 approaches offer optimized RNA targeting in zebrafish.
  • The study provides a refined CRISPR-RfxCas13d system with improved in vivo efficiency and reduced collateral activity.
  • Findings support the progression of CRISPR technology for in vivo RNA manipulation in zebrafish models.