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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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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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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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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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Efficient Targeted Mutagenesis in Brassica Crops Using CRISPR/Cas Systems.

Tom Lawrenson1, Mark Youles2, Monika Chhetry2

  • 1John Innes Centre, Norwich, Norfolk, UK. tom.lawrenson@JIC.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|March 30, 2023
PubMed
Summary

CRISPR gene editing offers improved efficiency for Brassica crops. This protocol details enhanced Cas9 and Cas12a systems for diverse plant mutagenesis, enabling more complex genetic modifications.

Keywords:
Brassica napusBrassica oleraceaCRISPR/Cas9Cas12aGenome editingTargeted mutagenesis

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Area of Science:

  • Plant molecular biology
  • Genetics and genomics
  • Agricultural biotechnology

Background:

  • CRISPR/Cas systems have been utilized for targeted mutagenesis in plants since 2013.
  • Applications include important crop species like Brassica napus and Brassica oleracea.
  • Ongoing research focuses on enhancing CRISPR system efficiency and expanding available tools.

Purpose of the Study:

  • To present an improved protocol for CRISPR/Cas9 gene editing in Brassica species.
  • To introduce and evaluate the Cas12a system as an alternative for plant genome editing.
  • To enable more challenging and diverse genetic modifications in Brassica crops.

Main Methods:

  • Utilizing an optimized CRISPR/Cas9 system for higher editing efficiency.
  • Implementing the CRISPR/Cas12a system for versatile DNA targeting.
  • Applying these systems to Brassica napus and Brassica oleracea for mutagenesis.

Main Results:

  • Demonstrated enhanced efficiency of the improved Cas9 system.
  • Successfully employed Cas12a for targeted mutagenesis, showcasing its utility.
  • Achieved diverse and complex editing outcomes in Brassica species.

Conclusions:

  • The presented protocol offers advanced CRISPR tools for Brassica crop improvement.
  • The combination of improved Cas9 and Cas12a broadens the scope of achievable genetic edits.
  • This work facilitates more sophisticated research and breeding applications in Brassica plants.