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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

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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

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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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Updated: Oct 15, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Protoplasts: From Isolation to CRISPR/Cas Genome Editing Application.

Jin-Jun Yue1, Jin-Ling Yuan1, Fu-Hui Wu2

  • 1Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou, China.

Frontiers in Genome Editing
|October 29, 2021
PubMed
Summary
This summary is machine-generated.

Protoplast technology enables efficient DNA-free gene editing using CRISPR/Cas systems in crops. This review highlights protoplast regeneration

Keywords:
CRISPR/cas (clustered regularly interspaced short palindromic repeats)DNA-freeRNPprotoplaststransient transfection

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Area of Science:

  • Plant biotechnology
  • Gene editing technologies
  • Crop improvement

Background:

  • Protoplasts are crucial for validating CRISPR/Cas system components like RNA-guided endonucleases and Cas proteins.
  • Protoplast technology facilitates DNA-free gene editing, particularly beneficial for crops with complex genetics or reproductive challenges.
  • Efficient protoplast regeneration is a critical bottleneck for successful DNA-free gene editing applications.

Purpose of the Study:

  • To review the historical development and future prospects of protoplast technology in plant breeding.
  • To explore various protoplast manipulation techniques, including transfection, transformation, fusion, and regeneration.
  • To summarize current applications of protoplast technology in CRISPR/Cas-based crop improvement.

Main Methods:

  • Review of existing literature on protoplast technology and CRISPR/Cas gene editing.
  • Analysis of protoplast applications in diverse crop species.
  • Discussion of advancements in protoplast regeneration and gene editing efficiency.

Main Results:

  • Protoplasts serve as a versatile platform for rapid assessment of CRISPR/Cas editing tools.
  • DNA-free gene editing via protoplasts is increasingly applied to recalcitrant crop species.
  • Protoplast regeneration protocols are continuously being optimized for broader crop applicability.

Conclusions:

  • Protoplast technology is integral to advancing DNA-free gene editing for crop breeding.
  • Future research should focus on enhancing protoplast regeneration and expanding its application scope.
  • CRISPR/Cas-mediated gene editing using protoplasts offers significant potential for accelerated crop development.