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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.1K
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

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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Related Experiment Video

Updated: Nov 26, 2025

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans
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CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans

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Gene Editing in Dimorphic Fungi Using CRISPR/Cas9.

Gregory C Kujoth1, Thomas D Sullivan1, Bruce S Klein1,2

  • 1Department of Pediatrics, University of Wisconsin-Madison, Madison, Wisconsin.

Current Protocols in Microbiology
|December 14, 2020
PubMed
Summary
This summary is machine-generated.

This study details using CRISPR/Cas9 gene editing in dimorphic fungi, specifically Blastomyces dermatitidis. This powerful technique enables precise genetic modifications for better understanding and treatment of fungal pathogens.

Keywords:
Agrobacterium-mediated transformationBlastomycesCRISPR/Cas9dimorphic fungigene targeting

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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Area of Science:

  • Mycology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Dimorphic fungi like Blastomyces, Histoplasma, Coccidioides, and Paracoccidioides are significant human pathogens globally.
  • Understanding their biology is crucial for developing new treatments and vaccines.
  • Gene editing technologies are vital research tools for these organisms.

Purpose of the Study:

  • To provide a detailed protocol for applying CRISPR/Cas9 gene editing technology to dimorphic fungi.
  • To use Blastomyces dermatitidis as a model organism for demonstrating these gene-editing techniques.
  • To guide researchers in designing and implementing gene-editing strategies for fungal pathogens.

Main Methods:

  • Design and construction of single-guide RNA and Cas9-expressing targeting vectors, including multiplexed options.
  • Introduction of plasmids into Blastomyces using Agrobacterium-mediated transformation.
  • Detailed protocols for vector construction, protospacer selection, and Agrobacterium/Blastomyces preparation.

Main Results:

  • Demonstration of CRISPR/Cas9 application in Blastomyces dermatitidis.
  • Guidance on expected gene-editing efficiency and the types of genetic alterations achievable.
  • Successful implementation of Agrobacterium-mediated transformation for gene targeting.

Conclusions:

  • CRISPR/Cas9 technology is a powerful and applicable tool for genetic manipulation in dimorphic fungi.
  • This protocol facilitates in-depth research into the biology of significant fungal pathogens.
  • The described methods will aid in the development of novel therapeutic strategies against fungal infections.