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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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Related Experiment Video

Updated: Apr 16, 2026

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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A CRISPR/Cas9 vector system for tissue-specific gene disruption in zebrafish.

Julien Ablain1, Ellen M Durand1, Song Yang1

  • 1Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana Farber Cancer Institute, Boston, MA 02115, USA.

Developmental Cell
|March 11, 2015
PubMed
Summary

Researchers developed a CRISPR-based vector for targeted gene silencing in zebrafish. This system enables tissue-specific gene knockout, specifically in red blood cells, aiding loss-of-function studies.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • CRISPR/Cas9 genome editing facilitates targeted gene inactivation.
  • Zebrafish are a key model organism for genetic studies.
  • Rapid generation of knockout lines is crucial for research.

Purpose of the Study:

  • To develop a simple and scalable CRISPR-based vector system for tissue-specific gene inactivation in zebrafish.
  • To demonstrate the utility of this system for studying gene function in specific cell lineages.

Main Methods:

  • Utilized a CRISPR-based vector system with a tissue-specific promoter (gata1) to drive Cas9 expression.
  • Targeted the urod gene, involved in heme biosynthesis, in the erythrocytic lineage.
  • Analyzed gene disruption and phenotypic consequences in F0 embryos and stable F1 zebrafish.

Main Results:

  • Achieved specific silencing of the urod gene in erythrocytic lineage using the gata1 promoter.
  • Observed red fluorescent erythrocytes in zebrafish embryos, mimicking the yquem mutant phenotype.
  • Demonstrated mosaic gene disruption in F0 generation, with highly penetrant phenotypes in F1 generation.

Conclusions:

  • The developed CRISPR vector system allows for spatially controlled gene knockout in zebrafish.
  • This tool significantly expands the possibilities for loss-of-function studies in zebrafish, particularly for tissue-specific gene functions.