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Related Concept Videos

CRISPR01:59

CRISPR

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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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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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Homologous Recombination02:31

Homologous Recombination

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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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Related Experiment Video

Updated: Dec 18, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

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Very fast CRISPR on demand.

Yang Liu1, Roger S Zou2, Shuaixin He3

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, MD, USA. tjha@jhu.edu bwu20@jhu.edu.

Science (New York, N.Y.)
|June 13, 2020
PubMed
Summary
This summary is machine-generated.

We developed very fast CRISPR (vfCRISPR), a light-activated genome editing tool. This method precisely controls DNA double-strand breaks for high-resolution studies of DNA repair dynamics.

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Related Experiment Videos

Last Updated: Dec 18, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas systems are powerful genome editing tools.
  • Precise temporal control over DNA cleavage is needed for detailed studies of DNA repair mechanisms.

Purpose of the Study:

  • To develop a light-inducible CRISPR-Cas9 system for high-resolution spatiotemporal analysis of DNA repair.
  • To investigate the kinetics and molecular events of DNA double-strand break repair.

Main Methods:

  • Developed a caged RNA strategy for light-activated Cas9 binding and cleavage (vfCRISPR).
  • Utilized single-cell fluorescence imaging to monitor DNA repair foci (e.g., 53BP1) and protein dynamics (e.g., MRE11).
  • Measured the propagation of H2AX phosphorylation as a marker of DNA damage response.

Main Results:

  • vfCRISPR enables rapid, synchronized DNA double-strand break induction at the second and submicrometer scales.
  • Observed cellular responses to DNA damage within minutes, including MRE11 retention post-ligation.
  • Characterized the kinetics of H2AX phosphorylation spread and 53BP1 repair foci dynamics.
  • Demonstrated imaging-guided, single-allele genomic manipulation.

Conclusions:

  • vfCRISPR provides unprecedented spatiotemporal resolution for studying DNA repair.
  • The system allows detailed kinetic analysis of DNA damage response pathways.
  • Enables precise genomic manipulation for advanced biological research.