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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

78
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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Homologous Recombination02:31

Homologous Recombination

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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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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: Jul 26, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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CRISPR/Cas9-based gene activation and base editing in Populus.

Tao Yao1,2, Guoliang Yuan1,2,3, Haiwei Lu1,4

  • 1Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.

Horticulture Research
|June 16, 2023
PubMed
Summary

This study demonstrates the effectiveness of CRISPR activation and base editing technologies in poplar trees. These advanced genome editing tools enable precise gene expression regulation and genetic engineering in woody species for improved traits.

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Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Forest Genetics

Background:

  • The genus *Populus* is vital for environmental, industrial, and biofuel applications, serving as a model for tree research.
  • CRISPR/Cas9 has been used for gene knockouts in *Populus*, but gene activation and base editing technologies remain largely unevaluated.
  • Genetic improvement of *Populus* is crucial for enhanced growth and tailored lignin composition.

Purpose of the Study:

  • To evaluate the efficacy of deactivated Cas9 (dCas9)-based CRISPR activation (CRISPRa) for gene expression fine-tuning in two *Populus* clones.
  • To assess the application of Cas9 nickase (nCas9)-based cytosine base editor (CBE) for precise gene engineering in *Populus*.
  • To demonstrate the utility of advanced CRISPR/Cas-based technologies for gene regulation and engineering in woody species.

Main Methods:

  • Employed a dCas9-based CRISPRa technique to modulate the expression of *TPX2* and *LecRLK-G* genes.
  • Utilized transient expression in protoplasts and *Agrobacterium*-mediated stable transformation for CRISPRa delivery.
  • Applied nCas9-based CBE to introduce C-to-T conversions in the *PLATZ* gene for premature stop codon induction.

Main Results:

  • CRISPRa successfully increased target gene expression by 1.2-fold to 7.0-fold in both transient and stable systems.
  • The Cas9 nickase-based cytosine base editor achieved 13%-14% efficiency in introducing C-to-T mutations in the *PLATZ* gene.
  • Demonstrated the effectiveness of dCas9-based CRISPRa and nCas9-based CBE in *Populus* species.

Conclusions:

  • Successfully applied CRISPR/Cas-based technologies for gene expression regulation and precise gene engineering in two *Populus* species.
  • The dCas9-based CRISPRa system is effective for fine-tuning gene expression in *Populus*.
  • Emerging genome editing tools like CRISPRa and CBE can be readily adopted for advancing research and applications in woody species.