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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

176
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...
176
CRISPR01:59

CRISPR

52.7K
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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Related Experiment Video

Updated: Aug 29, 2025

Author Spotlight: Streamlining Rice Breeding with CRISPR/Cas for Obtaining Optimal Phenotypic and Agronomic Traits
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A sequential transformation method for validating soybean genome editing by CRISPR/Cas9 system.

Duy Dinh Trinh1, Ngoc Thu Le1, Thao Phuong Bui1,2

  • 1Institute of Biotechnology, Vietnam Academy of Science and Technology, Hoang Quoc Viet, Cau Giay, Hanoi, Viet Nam.

Saudi Journal of Biological Sciences
|September 5, 2022
PubMed
Summary
This summary is machine-generated.

This study introduces a novel sequential transformation method for soybean genome editing using CRISPR/Cas9. The approach effectively validates guide RNA (gRNA) activity and assesses gene editing efficiency in soybean hairy roots.

Keywords:
Agrobacterium rhizogenesCRISPR/Cas9Guide RNAHairy rootsSequential transformationSoybean

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Area of Science:

  • Plant biotechnology
  • Molecular biology
  • Genome editing

Background:

  • CRISPR/Cas9 technology enables precise genome modifications.
  • Evaluating guide RNA (gRNA) efficacy is crucial for successful genome editing.
  • Soybean transformation systems are essential for genetic studies and crop improvement.

Purpose of the Study:

  • To evaluate a sequential transformation method for soybean genome editing using CRISPR/Cas9.
  • To demonstrate a strategy for assessing CRISPR/Cas9 construct activity, particularly guide RNAs (gRNAs).
  • To analyze the targeting efficacy of different gRNAs in soybean.

Main Methods:

  • Constructed gRNAs targeting exogenous (gus) and endogenous (SACPD-C, SMT) soybean genes.
  • Utilized Agrobacterium rhizogenes-mediated hairy root induction for soybean transformation.
  • Identified and characterized targeted mutations using poly-acrylamide gel electrophoresis (PAGE) heteroduplex method and sequencing.

Main Results:

  • Induced mutations in the gus gene in 57% of transgenic hairy roots.
  • Achieved 100% targeted mutation rate for the endogenous SACPD-C gene.
  • Observed mutation rates of 75% for SACPD-C and 67% for SMT using multiple gRNAs, with various indels detected.

Conclusions:

  • The sequential transformation method successfully validated gRNA targeting efficacy in soybean.
  • Discrepancies between bioinformatics predictions and experimental results for gRNA activity were identified.
  • This method represents the first application in soybean using a hairy root system, offering a valuable tool for CRISPR/Cas9 construct validation and gRNA evaluation.