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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.
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Conservative Site-specific Recombination and Phase Variation02:53

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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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Updated: Mar 30, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Combining CRISPR/Cas9 and rAAV Templates for Efficient Gene Editing.

Manuel Kaulich1, Steven F Dowdy1

  • 1Department of Cellular and Molecular Medicine, University of California , San Diego, La Jolla, California.

Nucleic Acid Therapeutics
|November 6, 2015
PubMed
Summary

CRISPR/Cas9 gene editing combined with recombinant adeno-associated virus (rAAV) significantly enhances gene targeting efficiency. This practical guide details applying this system for cost-effective genetic modification in cell biology research.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 is a key tool for altering endogenous genes.
  • CRISPR/Cas9 introduces specific genomic DNA breaks.
  • Non-homologous end joining (NHEJ) repairs breaks via deletions without a template.
  • Homology-directed repair (HDR) with a DNA template offers gene targeting but with variable efficiency.

Purpose of the Study:

  • To review the current state of CRISPR/Cas-based gene editing.
  • To provide a practical guide for efficient gene targeting using CRISPR/Cas and rAAV.
  • To highlight the time- and cost-effectiveness of this combined system.

Main Methods:

  • Utilizing CRISPR/Cas9 for targeted genomic DNA breaks.
  • Employing recombinant adeno-associated virus (rAAV) to deliver single-stranded homologous DNA templates.
  • Combining CRISPR/Cas9 and rAAV for homology-directed repair.

Main Results:

  • CRISPR/Cas9 combined with rAAV induces highly efficient gene targeting.
  • This method overcomes the variable efficiencies of traditional HDR.
  • The system allows for precise genomic alterations.

Conclusions:

  • The CRISPR/Cas9 and rAAV system represents a significant advancement in gene editing.
  • This approach enables highly efficient, time- and cost-effective gene targeting.
  • It offers a practical solution for researchers in cell biology and genetics.