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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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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

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Related Experiment Video

Updated: Apr 11, 2026

CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
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CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis

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Targeted mutagenesis in soybean using the CRISPR-Cas9 system.

Xianjun Sun1, Zheng Hu2, Rui Chen3

  • 11] College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China [2] National Key Facilities for Crop Genetic Resources and Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

Scientific Reports
|May 30, 2015
PubMed
Summary

CRISPR-Cas9 genome editing efficiently mutated target genes in soybean hairy roots. The soybean U6 promoter vector achieved higher mutation efficiencies, advancing soybean functional genomics research.

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High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots
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High-throughput CRISPR Vector Construction and Characterization of DNA Modifications by Generation of Tomato Hairy Roots

Published on: April 30, 2016

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • Genome editing is crucial for gene function analysis and crop improvement.
  • The CRISPR-Cas9 system offers precise gene editing capabilities.
  • Soybean (Glycine max L.) requires efficient tools for functional genomics.

Purpose of the Study:

  • To develop and evaluate CRISPR-Cas9 vectors for targeted gene mutagenesis in soybean.
  • To compare the efficiency of soybean and Arabidopsis U6 promoters in driving synthetic guide RNA (sgRNA) expression.
  • To assess gene editing efficiency and off-target effects in soybean hairy roots.

Main Methods:

  • Prediction of 11 U6 genes in soybean.
  • Construction of pCas9-GmU6-sgRNA and pCas9-AtU6-sgRNA vectors.
  • Transformation of vectors into soybean protoplasts and hairy roots via Agrobacterium rhizogenes.
  • Detection of mutations and off-target activities in target genes (Glyma06g14180, Glyma08g02290, Glyma12g37050).

Main Results:

  • Efficient target gene editing was achieved in soybean hairy roots.
  • The pCas9-GmU6-sgRNA vector showed higher mutation efficiencies (14.7-20.2%) compared to pCas9-AtU6-sgRNA (3.2-9.7%).
  • Biallelic mutations were detected, and off-target activities were identified, suggesting potential for library construction.

Conclusions:

  • CRISPR-Cas9 system with soybean U6 promoter is effective for soybean gene editing.
  • This approach advances functional genomic research in soybean, particularly for root and nodule genes.
  • Identified off-target mutations could be leveraged for creating saturated mutation libraries.