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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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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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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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Updated: May 6, 2026

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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Affordable CRISPR RNP-Based Genome Editing in Euglena gracilis.

Anzu Minami1,2, Minami Shimizu1,2, Shun Tamaki3

  • 1Center for Sustainable Resource Science, RIKEN, Yokohama, Kanagawa, Japan.

Current Protocols
|May 5, 2026
PubMed
Summary
This summary is machine-generated.

We developed an affordable and efficient genome editing workflow for Euglena gracilis using Cas9 nucleases. This method simplifies techniques, making gene editing more accessible for research and industrial applications in green algae.

Keywords:
CRISPR ribonucleoprotein (RNP)‐based genome editingCas9 nucleasesEuglena gracilistransformation

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Area of Science:

  • Algal biotechnology
  • Molecular biology
  • Synthetic biology

Background:

  • Genome editing in green algae like Euglena gracilis is crucial for advancing basic research and industrial applications.
  • Previous genome editing methods for Euglena gracilis were often technically demanding and required specialized equipment, limiting their widespread adoption.

Purpose of the Study:

  • To present an affordable and broadly applicable workflow for genome editing in Euglena gracilis.
  • To lower the technical barriers associated with genome editing in this organism.
  • To enable a wider range of genome editing outcomes, including deletions and base substitutions.

Main Methods:

  • Utilized Cas9 nucleases for genome editing in Euglena gracilis.
  • Employed a general-purpose laboratory electroporator for transformation.
  • Implemented a simplified clonal isolation procedure, avoiding specialized micromanipulation devices.
  • Detailed protocols for Euglena gracilis culture, sgRNA synthesis, transformation, and genotyping were established.

Main Results:

  • Achieved high genome editing efficiency in Euglena gracilis.
  • Demonstrated the compatibility of the workflow with various editing outcomes, such as targeted deletions and precise base substitutions.
  • Successfully adapted the protocol for use with standard laboratory equipment.

Conclusions:

  • The presented workflow significantly lowers technical barriers for genome editing in Euglena gracilis.
  • This accessible method facilitates broader application of genome editing for both fundamental research and industrial purposes in green algae.
  • The protocol supports diverse editing strategies, paving the way for enhanced genetic manipulation of Euglena gracilis.