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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

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

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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.
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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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Updated: Sep 22, 2025

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans
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Highly Efficient Genome Editing in Clostridium difficile Using the CRISPR-Cpf1 System.

Wei Hong1,2,3, Jie Zhang3, Guzhen Cui4

  • 1Key Laboratory of Endemic and Ethnic Diseases, Guizhou Medical University, Ministry of Education, Guiyang, Guizhou, China.

Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2022
PubMed
Summary
This summary is machine-generated.

We developed a new CRISPR-Cpf1 genome editing tool for Clostridium difficile, enhancing our ability to study its disease mechanisms. This efficient method aids in understanding and combating Clostridium difficile infections.

Keywords:
C. difficile infectionCRISPR-Cpf1Clostridium difficileGenome editing

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Clostridium difficile is a major cause of hospital-acquired diarrhea and mortality.
  • Efficient genome editing tools are needed to study C. difficile pathogenesis and physiology.
  • Current methods for genetic manipulation in C. difficile can be time-consuming.

Purpose of the Study:

  • To develop and validate a streamlined CRISPR-Cpf1-based genome editing protocol for C. difficile.
  • To establish a rapid DNA cloning method for plasmid construction.
  • To provide a versatile genome engineering tool for prokaryotic research.

Main Methods:

  • Application of the CRISPR-Cpf1 system for precise genome editing in C. difficile.
  • Development of a one-step-assembly protocol for DNA cloning.
  • Implementation of a Python-based algorithm for automated primer design.

Main Results:

  • Achieved high-efficiency, precise genome editing in C. difficile using CRISPR-Cpf1.
  • Demonstrated the first successful application of CRISPR-Cpf1 for C. difficile genome editing.
  • Reduced plasmid construction time by 50% compared to conventional methods.
  • Developed a broadly applicable genome engineering approach for microorganisms.

Conclusions:

  • The developed CRISPR-Cpf1 protocol is an efficient tool for C. difficile genome editing.
  • This technology is crucial for understanding C. difficile pathogenicity and developing infection control strategies.
  • The rapid cloning and primer design methods accelerate genetic research in prokaryotes.