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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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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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Updated: Sep 6, 2025

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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Engineering Citrobacter freundii using CRISPR/Cas9 system.

Trinidad Alfaro1, Joshua R Elmore2, Zachary R Stromberg1

  • 1Chemical and Biological Signatures Group, National Security Directorate, Pacific Northwest National Laboratory, P.O. Box 999, Richland, WA 99352, USA.

Journal of Microbiological Methods
|July 2, 2022
PubMed
Summary
This summary is machine-generated.

The CRISPR/Cas9 genome editing tool successfully modified the Citrobacter freundii bacterium, enabling efficient gene deletion and insertion. This expands the utility of CRISPR/Cas9 for diverse microbial engineering applications.

Keywords:
CRISPR/CasCitrobacter freundiiGenome editingsgRNA

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • CRISPR/Cas9 is a powerful genome editing tool, but its application in bacteria is limited.
  • Expanding CRISPR/Cas9 to new microbial hosts is crucial for broader biotechnological applications.

Purpose of the Study:

  • To evaluate the efficacy of the CRISPR/Cas9 system for genome editing in Citrobacter freundii.
  • To determine the efficiency of gene deletion and insertion in C. freundii using CRISPR/Cas9.

Main Methods:

  • Utilized the two-plasmid pCas/pTargetF system for CRISPR/Cas9-mediated genome editing.
  • Performed gene deletions (galactokinase) and insertions (mNeonGreen, yebA, xylS) in C. freundii ATCC 8090.
  • Assessed editing efficiency using various single-guide RNA sequences.

Main Results:

  • Achieved high editing efficiency (~91%) for galactokinase gene deletion.
  • Successfully inserted reporter and functional genes (mNeonGreen, yebA, xylS) with high efficiency (81-100%).
  • Demonstrated CRISPR/Cas9's effectiveness for both gene deletion and insertion in C. freundii.

Conclusions:

  • CRISPR/Cas9 is a suitable and efficient tool for genome editing in Citrobacter freundii.
  • This study expands the application of CRISPR/Cas9 technology to C. freundii, establishing it as a viable microbial chassis.