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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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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 and crRNAs02:53

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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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Homologous Recombination02:31

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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Fixing Double-strand Breaks02:04

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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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Updated: Aug 5, 2025

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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Mapping cellular responses to DNA double-strand breaks using CRISPR technologies.

Yang Liu1, W Taylor Cottle1, Taekjip Ha2

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, MD, USA.

Trends in Genetics : TIG
|March 26, 2023
PubMed
Summary
This summary is machine-generated.

This review explores using CRISPR-Cas9 as a tool to induce DNA double-strand breaks (DSBs) for studying their repair. Inducible Cas9 systems combined with advanced imaging and sequencing offer new mechanistic insights into cellular responses to DNA damage.

Keywords:
CRISPR-Cas9DNA double-strand breakDSB repairchromatingenomic imagingvery fast CRISPR

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA double-strand breaks (DSBs) are highly genotoxic lesions linked to cancer and immunodeficiency.
  • Cells possess complex DNA repair networks to maintain genomic stability against DSBs.
  • CRISPR-Cas9 technology has renewed interest in studying DSB repair mechanisms.

Purpose of the Study:

  • To review advancements in using Cas9 as a precise inducer of DSBs for repair studies.
  • To highlight how Cas9 facilitates the investigation of cellular responses to DNA damage.
  • To showcase novel insights into DSB repair mechanisms enabled by Cas9 technology.

Main Methods:

  • Utilizing Cas9 as a targeted DNA damage inducer.
  • Employing rapidly inducible Cas9 systems for synchronized break generation.
  • Integrating sequencing and genome-specific imaging with Cas9-induced DSBs.

Main Results:

  • Inducible Cas9 systems allow for controlled and efficient induction of DSBs.
  • Combined approaches enable spatiotemporal characterization of cellular DSB responses.
  • Mechanistic insights into DSB repair at specific genomic locations are now achievable.

Conclusions:

  • Cas9 is a powerful tool for dissecting DNA double-strand break repair pathways.
  • Advanced methodologies enhance our ability to study cellular responses to DNA damage.
  • This approach provides unprecedented mechanistic understanding of genomic instability and repair.