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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

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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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Homologous Recombination02:31

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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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Related Experiment Video

Updated: Dec 18, 2025

Author Spotlight: Streamlining Rice Breeding with CRISPR/Cas for Obtaining Optimal Phenotypic and Agronomic Traits
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Author Spotlight: Streamlining Rice Breeding with CRISPR/Cas for Obtaining Optimal Phenotypic and Agronomic Traits

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CRISPR/Cas9-Based Genome Editing Using Rice Zygotes.

Erika Toda1, Takashi Okamoto1

  • 1Department of Biological Sciences, Tokyo Metropolitan University, Hachioji, Tokyo, Japan.

Current Protocols in Plant Biology
|June 10, 2020
PubMed
Summary

This study presents a new CRISPR/Cas9 genome-editing protocol for rice zygotes, overcoming delivery and transformation hurdles. This method enables efficient targeted mutagenesis for advanced plant breeding and crop improvement.

Keywords:
CRISPR/Cas9PEG-Ca2+ transfectiongenome editingribonucleoproteinricezygote

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Agricultural Science

Background:

  • Genome editing offers significant potential for next-generation plant breeding.
  • Existing applications are often limited by challenges in macromolecule delivery, plant transformation, and regeneration.

Purpose of the Study:

  • To describe a novel protocol for CRISPR/Cas9-based genome editing in rice zygotes.
  • To overcome technical barriers in plant genome editing for efficient crop improvement.

Main Methods:

  • Rice zygotes were produced via in vitro fertilization of isolated rice gametes.
  • CRISPR/Cas9 components (plasmid DNA or ribonucleoproteins) were delivered into zygotes using polyethylene glycol/calcium-mediated transfection.
  • Transfected zygotes were regenerated into whole plants.

Main Results:

  • The protocol achieved high-frequency targeted mutagenesis in regenerated rice plants.
  • Mutation frequencies ranged from mono-allelic to bi-allelic, between 4% and 64%.
  • Successful regeneration of edited plants was achieved.

Conclusions:

  • This method provides an efficient protocol for CRISPR/Cas9 genome editing in rice zygotes.
  • The protocol has the potential to significantly advance molecular breeding in rice and other crop species.
  • Overcoming delivery and transformation hurdles facilitates broader application of genome editing in agriculture.