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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: Nov 20, 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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Efficient Genome Editing in Rice Protoplasts Using CRISPR/CAS9 Construct.

Martine Bes1,2, Leo Herbert1,2, Thibault Mounier1,2

  • 1CIRAD, UMR AGAP, Montpellier Cedex 5, France.

Methods in Molecular Biology (Clifton, N.J.)
|January 20, 2021
PubMed
Summary
This summary is machine-generated.

This study presents a new, efficient protoplast-based protocol for testing CRISPR/Cas9 genome editing technologies in rice plants. This method significantly speeds up the evaluation of new gene editing tools, reducing time and labor for plant breeding applications.

Keywords:
CRISPR/CAS9Genome editingProtoplastRice

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Area of Science:

  • Plant Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 and related genome editing technologies are transforming plant biology and breeding.
  • Evaluating new editing technologies in whole plants is time-consuming and labor-intensive, often requiring transgenic plant production.
  • There is a need for efficient, high-throughput methods to assess genome editing tool efficacy in plants.

Purpose of the Study:

  • To develop a routine, high-throughput protocol for testing genome editing technologies in rice using protoplasts.
  • To establish a rapid system for evaluating the effectiveness of CRISPR/Cas9 constructs in plant cells.
  • To provide a comprehensive protocol from sgRNA design to mutation analysis.

Main Methods:

  • Utilization of rice protoplasts (plant cells lacking cell walls) as a model system.
  • Development of a protocol to test over 30 constructs in protoplasts derived from 9-11-day-old seedlings.
  • Application of CRISPR/Cas9 technology for genome editing assessments.

Main Results:

  • The developed protocol allows for efficient testing of genome editing technologies in rice protoplasts.
  • CRISPR/Cas9 construct effectiveness can be determined within a week.
  • The protocol enables high-throughput screening of multiple gene editing constructs.

Conclusions:

  • Protoplast-based systems offer a rapid and efficient alternative for evaluating plant genome editing technologies.
  • This protocol streamlines the assessment of CRISPR/Cas9 efficiency in rice, accelerating plant breeding research.
  • The findings facilitate the broader adoption and development of advanced genome editing tools in agriculture.