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Related Concept Videos

CRISPR01:59

CRISPR

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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: Jun 28, 2025

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Genome Editing in Brassica juncea Using CRISPR/Cas9 Technology.

Niaz Ahmad1, Samia Fatima2, Penny Hundleby3

  • 1National Institute for Biotechnology and Genetic Engineering College (NIBGE-C), Pakistan Institute for Engineering and Applied Sciences (PIEAS), Faisalabad, Pakistan. nahmad@nibge.org.

Methods in Molecular Biology (Clifton, N.J.)
|April 24, 2024
PubMed
Summary

This study presents a CRISPR/Cas9 genome editing protocol for Brassica juncea, a vital oilseed crop. The method enables precise genetic modifications to enhance crop resilience and facilitate functional genomic studies in this important field crop.

Keywords:
B. junceaCRISPR/Cas9KnockoutMultiplexingPolyploidTransgene free

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Area of Science:

  • Plant Biotechnology
  • Genomics
  • Crop Science

Background:

  • CRISPR/Cas9 technology has transformed plant genome manipulation, enabling enhanced resilience to climate change and diseases.
  • Current applications are largely confined to model crops, limiting broader agricultural benefits.
  • Polyploid species present unique challenges for genome editing due to multiple genome sets.

Purpose of the Study:

  • To establish a robust CRISPR/Cas9-based genome editing protocol for the allotetraploid crop Brassica juncea.
  • To enable precise genetic engineering for improved crop traits and functional genomic analysis in a key oilseed crop.
  • To extend advanced genome editing applications beyond model plants to important field crops.

Main Methods:

  • Agrobacterium-mediated transformation was employed for delivering CRISPR components.
  • Cotyledon explants were utilized for efficient transformation and regeneration.
  • A segregating approach was implemented for the recovery of transgene-free edited plants.

Main Results:

  • A detailed protocol for genome editing in Brassica juncea was successfully developed and validated.
  • The method allows for the recovery of genome-edited knockouts in this important crop.
  • Transgene-free edited plants were efficiently recovered, crucial for regulatory and commercial applications.

Conclusions:

  • The developed protocol significantly advances the application of CRISPR/Cas9 technology in Brassica juncea.
  • This methodology provides a powerful tool for accelerating crop improvement and functional genomics in an important oilseed crop.
  • Extending precise genome editing to field crops like Brassica juncea is vital for addressing agricultural challenges posed by climate change and disease.