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Related Concept Videos

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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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Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system.

Uwe Irion1, Jana Krauss2, Christiane Nüsslein-Volhard2

  • 1Max-Planck-Institut für Entwicklungsbiologie, Spemannstr. 35, Tübingen 72076, Germany uwe.irion@tuebingen.mpg.de.

Development (Cambridge, England)
|November 21, 2014
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Summary

CRISPR/Cas gene editing precisely repaired a mutation in zebrafish, enabling germline transmission of the corrected gene. This demonstrates zebrafish

Keywords:
CRISPR/CasGenome editingZebrafishalbinoslc45a2

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Area of Science:

  • Molecular Biology
  • Genetics
  • Zebrafish Models

Background:

  • Engineered nucleases enable precise genome editing.
  • Zebrafish research has achieved loss-of-function alleles but not germline transmission of targeted knock-ins or SNP exchanges.

Purpose of the Study:

  • To demonstrate CRISPR/Cas system efficacy in targeting and repairing a specific mutation in zebrafish.
  • To achieve germline transmission of a precise single nucleotide polymorphism (SNP) exchange in zebrafish.

Main Methods:

  • Utilized the CRISPR/Cas system for genome editing.
  • Employed circular donor DNA with CRISPR target sites for homology-directed repair.
  • Phenotypic rescue was used to assess repair efficiency.

Main Results:

  • Achieved high efficiency and precision in targeting and repairing a premature stop codon at the albino (alb) locus.
  • Nearly 50% of larvae showed precise homology-directed repair of the alb(b4) mutation.
  • Demonstrated germline transmission of the repaired allele in a small fraction of adult fish (3/28).

Conclusions:

  • The CRISPR/Cas system effectively targets and repairs mutations in zebrafish.
  • Successful in vivo demonstration of germline transmission of a precise SNP exchange.
  • Highlights zebrafish as a suitable model for genetic research and genome editing studies.