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Related Concept Videos

CRISPR01:59

CRISPR

49.3K
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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Optimization of a hypercompact Fanzor2 system for improved genome editing performance in plants.

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Coupling of both a transactivation module and a double-stranded DNA-binding domain boosts Cas12i3 variant-based cytosine and adenine editing in plants.

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Analysis of a Series of Mutants Highlights Complex Regulation of Fusarium Head Blight Resistance Conferred by Fhb1 Locus in Wheat.

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Functional conservation and divergence of the WOX gene family in regulating meristem activity: From Arabidopsis to crops.

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Fusion of a rice endogenous <i>N</i>-methylpurine DNA glycosylase to a plant adenine base transition editor ABE8e enables A-to-K base editing in rice plants.

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Engineering a robust Cas12i3 variant-mediated wheat genome editing system.

Wenxue Wang1,2, Lei Yan1, Jingying Li1,2

  • 1State Key Laboratory of Crop Gene Resources and Breeding, Institute of Crop Sciences (ICS), Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.

Plant Biotechnology Journal
|December 18, 2024
PubMed
Summary

Researchers developed a new CRISPR/Cas12i3 genome editing system for wheat. This enhanced system, Opt-T5E-Cas12i3-5M, significantly improves editing efficiency in wheat, aiding crop improvement.

Keywords:
CRISPR/Cas12i3T5 Exonuclease (T5E)genome editingwheat (Triticum aestivum L.)

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Area of Science:

  • Plant Biotechnology
  • Genome Editing
  • Crop Science

Background:

  • Wheat (Triticum aestivum L.) is a globally vital food crop.
  • CRISPR/Cas12i3 offers advantages like smaller protein size and flexible PAM recognition.
  • Previous attempts at Cas12i3-mediated editing in wheat were limited by low efficiency and the crop's hexaploid nature.

Purpose of the Study:

  • To engineer a robust Cas12i3-5M genome editing system for wheat.
  • To enhance editing efficiency in hexaploid wheat through system optimization.

Main Methods:

  • Fusion of T5 exonuclease (T5E) to Cas12i3-5M.
  • Optimization of crRNA expression strategy (Opt).
  • Testing the Opt-T5E-Cas12i3-5M system on four endogenous genes in three elite Chinese wheat varieties.

Main Results:

  • T5E fusion increased Cas12i3-5M editing efficiency in HEK293T cells by up to 3.87-fold.
  • The optimized system (Opt-T5E-Cas12i3-5M) achieved 1.20- to 7.95-fold higher editing efficiency in wheat stable lines compared to previous versions.
  • Editing efficiencies ranged from 60.71% to 90.00% across target genes in stable wheat lines.

Conclusions:

  • The developed Opt-T5E-Cas12i3-5M system is a robust tool for wheat genome editing.
  • This system enhances editing efficiency in polyploid wheat, overcoming previous limitations.
  • The technology holds potential for accelerating wheat research and genetic improvement, and may be applicable to other polyploid crops.