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Related Concept Videos

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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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...
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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Genome Editing in Astyanax mexicanus Using Transcription Activator-like Effector Nucleases (TALENs)
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In vivo genome editing using a high-efficiency TALEN system.

Victoria M Bedell1, Ying Wang, Jarryd M Campbell

  • 1Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA.

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Enhanced artificial transcription activator-like effector nucleases (TALENs) enable precise genome editing in zebrafish. This breakthrough facilitates targeted gene modification and functional genomic studies in this important vertebrate model.

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Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Area of Science:

  • Molecular Biology
  • Genetics
  • Zebrafish Model Systems

Background:

  • Zebrafish (Danio rerio) are vital for studying vertebrate biology and human diseases.
  • Previous limitations in targeted genome modification hindered zebrafish research.
  • Artificial transcription activator-like effector nucleases (TALENs) offer potential for genome editing.

Purpose of the Study:

  • To evaluate enhanced TALEN technology for targeted zebrafish genome editing.
  • To demonstrate the efficiency of TALENs in inducing locus-specific DNA breaks.
  • To showcase precise sequence modification via homology-directed repair in zebrafish.

Main Methods:

  • Utilized an improved GoldyTALEN scaffold and zebrafish delivery system.
  • Administered TALENs to induce DNA breaks in somatic and germline tissues.
  • Employed single-stranded DNA oligonucleotides for homology-directed repair.

Main Results:

  • Achieved high-efficiency, locus-specific DNA breaks with TALENs, approaching 100% at some loci.
  • Demonstrated biallelic conversion in somatic tissues, mimicking morpholino knockdown phenotypes.
  • Successfully introduced custom sequences (EcoRV site, mloxP) and achieved germline transmission.

Conclusions:

  • Enhanced TALENs provide a powerful tool for targeted zebrafish genome editing.
  • This method enables precise sequence modification and functional genomic applications.
  • The approach facilitates modeling genetic variation and generating targeted conditional alleles in zebrafish.