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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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CRISPR01:59

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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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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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Related Experiment Video

Updated: Mar 24, 2026

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
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2C-Cas9: a versatile tool for clonal analysis of gene function.

Vincenzo Di Donato1, Flavia De Santis1, Thomas O Auer1

  • 1Institut Curie, PSL Research University, INSERM U 934, CNRS UMR3215, F-75005, Paris, France;

Genome Research
|March 10, 2016
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 gene editing in zebrafish can now achieve tissue-specific gene inactivation. This new method allows for precise control and genetic labeling of mutant cell clones for detailed analysis.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • CRISPR/Cas9 is a powerful tool for creating loss-of-function mutations in zebrafish.
  • Current methods primarily generate constitutive knockouts.
  • Studying gene function often requires controlled inactivation at specific times and locations.

Purpose of the Study:

  • To develop a method for spatiotemporal control of gene inactivation in zebrafish using CRISPR/Cas9.
  • To establish a system for genetically labeling and analyzing mutant cell clones.
  • To provide a versatile tool for functional genomics in model organisms.

Main Methods:

  • Utilized the Gal4/UAS system to drive Cas9 expression in a tissue-specific manner.
  • Combined Gal4/UAS with the Cre/loxP system for inducible gene disruption and clonal labeling.
  • Applied CRISPR/Cas9 technology for targeted mutagenesis in zebrafish.

Main Results:

  • Demonstrated successful tissue-specific gene disruption by controlling Cas9 expression.
  • Established a dual system (Gal4/UAS and Cre/loxP) for generating and labeling mutant clones.
  • Enabled phenotypic analysis of genetically labeled mutant cell populations.

Conclusions:

  • The developed CRISPR/Cas9 system offers precise spatiotemporal control over gene inactivation in zebrafish.
  • This technique facilitates the study of gene function in specific tissues and cell types.
  • The method is adaptable to various model organisms for advanced functional genetic studies.