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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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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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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Efficient CRISPR/Cas9-Mediated Genome Editing Using a Chimeric Single-Guide RNA Molecule.

Haroon Butt1, Ayman Eid1, Zahir Ali1

  • 1Laboratory for Genome Engineering, Division of Biological Sciences, King Abdullah University of Science and TechnologyThuwal, Saudi Arabia.

Frontiers in Plant Science
|September 9, 2017
PubMed
Summary
This summary is machine-generated.

This study introduces a novel CRISPR/Cas9 method using chimeric guide RNA (cgRNA) for efficient gene editing in rice. The cgRNA system streamlines homology-directed repair (HDR) for improved crop trait development.

Keywords:
CRISPR/Cas9HDRRNA-templated repairgene editinggenome engineering

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • CRISPR/Cas9 enables targeted mutagenesis in eukaryotes.
  • Homology-directed repair (HDR) for precise gene editing often requires simultaneous delivery of DNA templates, limiting efficiency.

Purpose of the Study:

  • To develop a more efficient CRISPR/Cas9-based gene editing system in rice (Oryza sativa).
  • To utilize chimeric single-guide RNA (cgRNA) for simultaneous double-strand break induction and homology-directed repair (HDR) template delivery.

Main Methods:

  • Employed CRISPR/Cas9 technology with chimeric single-guide RNA (cgRNA) molecules in rice protoplasts.
  • cgRNAs integrated target site recognition and repair template sequences for HDR.
  • Investigated the efficiency of repair templates complementary to either the target or non-target DNA strand.

Main Results:

  • Achieved more efficient gene editing in rice protoplasts using repair templates complementary to the non-target DNA strand.
  • Successfully generated herbicide resistance in rice using the cgRNA-mediated HDR method.
  • Demonstrated the applicability of this cgRNA repair strategy for targeted gene editing in plants.

Conclusions:

  • The developed cgRNA repair method enhances CRISPR/Cas9-mediated gene editing efficiency in rice.
  • This approach facilitates targeted genetic improvement of crop traits and advances functional genomics in plants.