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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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CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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A simple and accessible CRISPR genome editing laboratory exercise using yeast.

Connor Shortt1, Elise Krippaehne1, Brian M Wasko1

  • 1College of Osteopathic Medicine of the Pacific Northwest, Western University of Health Sciences, Lebanon, Oregon, USA.

Micropublication Biology
|February 24, 2023
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Summary

This study introduces a CRISPR gene editing lab for students. Yeast cells edited to disrupt the ADE2 gene turn red, demonstrating genome engineering principles.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR technology offers precise genome engineering capabilities.
  • Educational tools are needed to teach CRISPR principles effectively.
  • Yeast genetics provides a tractable model for gene editing studies.

Purpose of the Study:

  • To develop an accessible CRISPR laboratory exercise for students.
  • To demonstrate genome editing by disrupting the ADE2 gene in yeast.
  • To enable students to observe phenotypic changes resulting from gene editing.

Main Methods:

  • Utilized CRISPR-Cas9 system for targeted genome editing in yeast.
  • Disrupted the ADE2 gene, a key gene in adenine biosynthesis.
  • Cultured yeast on selective media to identify edited colonies.

Main Results:

  • Successfully generated yeast colonies with disrupted ADE2 genes.
  • Edited yeast colonies exhibited a distinct red color phenotype.
  • The experiment was completed within a single laboratory session.

Conclusions:

  • The developed CRISPR exercise effectively introduces genome editing concepts.
  • Yeast ADE2 gene disruption is a suitable model for demonstrating CRISPR applications.
  • This laboratory exercise is accessible to beginners in yeast genetics and CRISPR technology.