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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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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

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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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Related Experiment Video

Updated: Sep 22, 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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Boosting plant genome editing with a versatile CRISPR-Combo system.

Changtian Pan1, Gen Li1, Aimee A Malzahn1

  • 1Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.

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|May 20, 2022
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Summary

We developed CRISPR-Combo, a versatile platform for simultaneous plant genome editing and gene activation. This tool accelerates crop breeding by enhancing genome engineering efficiency and enabling faster screening of edited plants.

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Area of Science:

  • Plant Biotechnology
  • Genome Engineering
  • Molecular Biology

Background:

  • CRISPR-Cas9, base editors, and CRISPR activation systems are crucial for plant genome engineering.
  • Current applications often use these systems separately, limiting their combined potential.
  • A unified system for simultaneous editing and activation is needed to advance plant science.

Purpose of the Study:

  • To develop a versatile CRISPR-Combo platform for simultaneous genome editing and gene activation in plants.
  • To demonstrate the platform's utility in accelerating plant genome engineering and crop improvement.
  • To showcase applications in Arabidopsis, poplar, and rice for enhanced breeding strategies.

Main Methods:

  • Development of a single Cas9 protein-based CRISPR-Combo platform.
  • Simultaneous targeted mutagenesis or base editing and gene activation.
  • Application in model plants (Arabidopsis, poplar) and a key crop (rice).

Main Results:

  • CRISPR-Combo shortened the Arabidopsis life cycle by activating a florigen gene, simplifying screening.
  • Accelerated regeneration and propagation of genome-edited poplar plants via activation of morphogenic genes.
  • Achieved hormone-independent rice regeneration, enriching heritable targeted mutations.

Conclusions:

  • CRISPR-Combo is a versatile tool for simultaneous plant genome editing and gene activation.
  • The platform significantly boosts plant genome engineering efficiency.
  • CRISPR-Combo shows promising applications for accelerating crop breeding and improving plant traits.