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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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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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Updated: Feb 22, 2026

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus.

Qi Liu1,2, Yu Jiang3,4, Lei Shao2

  • 1School of Pharmacy, Fudan University, Shanghai 201203, China.

Acta Biochimica Et Biophysica Sinica
|September 16, 2017
PubMed
Summary

We developed a novel CRISPR/Cas9 genome editing tool for Staphylococcus aureus. This system enables rapid, scarless genetic manipulation, accelerating the study of this important pathogenic bacterium.

Keywords:
Staphylococcigenegenotypingmolecular geneticsplasmid

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Genetics

Background:

  • Staphylococcus aureus is a major cause of nosocomial infections, characterized by high pathogenicity and multi-drug resistance.
  • Existing genetic manipulation tools for S. aureus have limitations, including genomic scarring, low efficiency, and lengthy experimental timelines.

Purpose of the Study:

  • To develop a rapid, efficient, and scarless genome editing tool for Staphylococcus aureus using a single-plasmid CRISPR/Cas9 system.

Main Methods:

  • A single-plasmid CRISPR/Cas9 system was constructed, incorporating the cas9 gene, a single guide RNA, and donor DNA.
  • The system utilizes a constitutive promoter (Pxyl/tet) for cas9 expression and a strong promoter (Pspac) for sgRNA transcription.
  • The vector was validated by performing gene deletions (tgt, rocA) and gene insertion (ermR cassette) in S. aureus.

Main Results:

  • Demonstrated successful and efficient chromosomal manipulation in S. aureus.
  • Achieved marker-free and scarless genetic modifications, including gene deletion and insertion.
  • The developed CRISPR/Cas9 tool significantly reduces the time required for genetic manipulation.

Conclusions:

  • Established a versatile CRISPR/Cas9 genome editing platform for S. aureus.
  • This tool overcomes limitations of previous methods, enabling faster and more precise genetic studies.
  • Accelerates research into the molecular mechanisms and pathogenicity of Staphylococcus aureus.