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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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Related Experiment Video

Updated: Nov 20, 2025

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans
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Targeted Genetic Changes in Candida albicans Using Transient CRISPR-Cas9 Expression.

Manning Y Huang1, Max C Cravener2, Aaron P Mitchell2

  • 1Department of Biochemistry and Biophysics, University of California San Francisco School of Medicine, San Francisco, California.

Current Protocols
|January 25, 2021
PubMed
Summary

This study introduces CRISPR-Cas9 technology for efficient gene modification in Candida albicans. It enables rapid generation of homozygous mutants, aiding virulence factor research.

Keywords:
CRISPR-Cas9Candida albicansgene deletiongene editingtransient CRISPR-Cas9 expression

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Mycology

Background:

  • Candida albicans is a significant opportunistic fungal pathogen causing disease and mortality.
  • Studying its virulence factors is crucial but challenging due to its diploid nature and lack of simple genetic manipulation tools.
  • Existing methods for gene modification are often complex and time-consuming.

Purpose of the Study:

  • To develop and present a robust CRISPR-associated nuclease 9 (Cas9) system for efficient gene modification in Candida albicans.
  • To enable the generation of homozygous mutants in a single transformation step.
  • To provide a versatile tool for dissecting Candida albicans virulence factors.

Main Methods:

  • Transient expression of CRISPR-Cas9 and single-guide RNAs from PCR cassettes.
  • Design principles and practical protocols for component amplification and Candida albicans transformation.
  • Methods for selecting and genotyping transformants, including CRISPR-induced marker excision (CRIME) for recyclable markers and knock-in strategies.

Main Results:

  • Successful implementation of CRISPR-Cas9 for gene modification in Candida albicans.
  • Generation of homozygous mutants in a single transformation step, significantly streamlining the process.
  • Demonstrated utility for gene deletion, knock-in, and combining multiple genetic modifications.

Conclusions:

  • The described CRISPR-Cas9 system offers a powerful and efficient approach for genetic manipulation of Candida albicans.
  • This methodology facilitates the study of gene function and virulence factors in this important fungal pathogen.
  • The protocols provide a valuable resource for researchers working with Candida albicans.