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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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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: Feb 22, 2026

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 genome editing in wheat.

Dongjin Kim1, Burcu Alptekin1, Hikmet Budak2

  • 1Cereal Genomics Lab, Department of Plant Sciences and Plant Pathology, Montana State University, Bozeman, MT, USA.

Functional & Integrative Genomics
|September 18, 2017
PubMed
Summary
This summary is machine-generated.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system effectively edited stress-responsive genes in wheat protoplasts. This demonstrates the system

Keywords:
CRISPR/Cas9Genome editingTaDREB2TaERF3Wheat

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Area of Science:

  • Plant Molecular Biology
  • Genome Editing Technologies
  • Crop Science

Background:

  • Genome editing in plants with complex genomes, like wheat, presents significant challenges.
  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system offers versatile gene editing capabilities guided by small RNAs.
  • While effective in diploid plants, CRISPR/Cas9 application in polyploid species remains challenging.

Purpose of the Study:

  • To apply the CRISPR/Cas9 genome editing system in wheat protoplasts.
  • To achieve targeted editing of stress-responsive transcription factor genes: wheat dehydration responsive element binding protein 2 (TaDREB2) and wheat ethylene responsive factor 3 (TaERF3).

Main Methods:

  • Transient expression of small guide RNA and Cas9 protein in wheat protoplasts.
  • Confirmation of mutagenesis using restriction enzyme digestion assay, T7 endonuclease assay, and sequencing.
  • Analysis of off-target regions and confirmation of genome editing specificity via amplicon sequencing.

Main Results:

  • Targeted genome editing of TaDREB2 and TaERF3 was successfully achieved in wheat protoplasts.
  • Mutagenesis effectiveness was confirmed through multiple molecular assays.
  • High specificity of the CRISPR/Cas9 system was demonstrated with minimal off-target mutations.

Conclusions:

  • The CRISPR/Cas9 genome editing system is readily applicable to wheat protoplasts.
  • This technology holds significant potential for targeted genetic manipulation and crop improvement in wheat.