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Related Concept Videos

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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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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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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Overview of Transposition and Recombination02:13

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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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.
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Related Experiment Video

Updated: Jul 28, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Characteristics, Recombination Methods, and Applications Progresses of Split-Cas9 System.

Zhixi Liu1,2, Lu Huang1,2, Han Deng1,2

  • 1Department of Pharmacy, Sichuan Clinical Research Center for Cancer, Sichuan Cancer Hospital & Institute, Sichuan Cancer Center, Affiliated Cancer Hospital of University of Electronic Science and Technology of China, Chengdu, China.

Human Gene Therapy
|June 2, 2023
PubMed
Summary

A novel Split-Cas9 system enhances CRISPR gene editing by dividing the Cas9 protein. This innovation improves therapeutic applications for genetic disorders and cancers by increasing vector compatibility and control.

Keywords:
CRISPRdelivery methodgene editinginteinsplit-Cas9 system

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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Last Updated: Jul 28, 2025

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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Editing

Background:

  • CRISPR technology offers potent gene editing capabilities with potential applications in treating genetic disorders and cancers.
  • Current CRISPR systems face challenges in delivery and precise control for therapeutic use.

Purpose of the Study:

  • To introduce and examine a novel Split-Cas9 system for advanced CRISPR gene editing.
  • To evaluate the combination modes, split sites, and activity efficiency of the Split-Cas9 system.
  • To discuss the potential clinical transformation of Split-Cas9 for in vivo and in vitro applications.

Main Methods:

  • The study involves splitting the Cas9 protein into fragments that recombine within cells under specific conditions.
  • Analysis of different Split-Cas9 combination strategies and their impact on gene editing efficiency.
  • Comparative assessment of various split sites within the Cas9 protein.

Main Results:

  • The Split-Cas9 system demonstrates improved compatibility with viral vectors due to smaller fragment sizes.
  • Precise temporal and spatial control over gene editing is achieved through the Split-Cas9 system.
  • Variations in split sites affect the activity efficiency of the Split-Cas9 system.

Conclusions:

  • The Split-Cas9 system offers a promising advancement in CRISPR technology, enhancing its therapeutic index.
  • This system provides improved delivery and control, paving the way for safer and more effective gene therapies.
  • Further research and clinical translation of Split-Cas9 hold significant potential for treating genetic diseases.