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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

142
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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What is Genetic Engineering?00:49

What is Genetic Engineering?

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Overview
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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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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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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.
The recognition sites for Cre recombinase called LoxP...
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Updated: Aug 23, 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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Frankenstein Cas9: engineering improved gene editing systems.

Pascal D Vos1,2,3,4,5, Aleksandra Filipovska3,4,5,6, Oliver Rackham1,2,3,4,6

  • 1Curtin Medical School, Curtin University, Bentley, Western Australia 6102, Australia.

Biochemical Society Transactions
|October 28, 2022
PubMed
Summary
This summary is machine-generated.

CRISPR-Cas9 gene editing is powerful but has limitations. Protein engineering has created improved Cas9 variants, offering enhanced tools for biotechnology and expanding the scope of CRISPR applications.

Keywords:
CRISPRdesigner Cas9enzyme designgene editingprotein engineeringsynthetic biology

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genetic Engineering

Background:

  • The CRISPR-Cas9 system has revolutionized biological research due to its targeted gene editing capabilities.
  • Guide RNA (gRNA) enables specific targeting of Cas9 nuclease activity.
  • Current Cas9 systems face challenges including off-target effects and variable on-target efficiency.

Purpose of the Study:

  • To address limitations of the standard CRISPR-Cas9 system.
  • To explore protein engineering strategies for improving Cas9 functionality.
  • To develop enhanced Cas9 variants for biotechnological applications.

Main Methods:

  • Protein engineering of the Cas9 nuclease.
  • Development of novel guide RNA designs.
  • Assessment of Cas9 variant activity, specificity, and fidelity.

Main Results:

  • Several Cas9 variants with improved nuclease activity have been engineered.
  • Variants demonstrate reduced off-target effects and enhanced on-target efficiency.
  • New Cas9 variants offer alternative functionalities for gene editing.

Conclusions:

  • Engineered Cas9 variants represent significant advancements over the original system.
  • These improved tools expand the potential and precision of CRISPR-based technologies.
  • Future biotechnological and therapeutic applications will benefit from these enhanced gene editing tools.