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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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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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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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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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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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Guide RNA engineering for versatile Cas9 functionality.

Chance M Nowak1,2, Seth Lawson1, Megan Zerez1,2

  • 1Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX 75080, USA.

Nucleic Acids Research
|October 14, 2016
PubMed
Summary
This summary is machine-generated.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system uses a single guide RNA (sgRNA) to direct Cas9 protein for genome editing. This review highlights sgRNA engineering advances for improved Cas9-mediated DNA targeting.

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Last Updated: Mar 13, 2026

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system, particularly Streptococcus pyogenes Cas9 (SpCas9), has transformed genome research and editing.
  • The system's effectiveness relies on the single guide RNA (sgRNA) directing the Cas9 protein to specific DNA sequences.

Purpose of the Study:

  • This review focuses on the properties and recent engineering advancements of the sgRNA component in Cas9-mediated genome targeting.
  • To explore how sgRNA modifications impact Cas9 association and function in genome editing.

Main Methods:

  • Literature review of CRISPR-Cas9 system research.
  • Analysis of studies detailing sgRNA structure, function, and modifications.
  • Examination of engineering strategies for enhancing sgRNA programmability and tolerance.

Main Results:

  • Cas9 protein's modularity is significantly influenced by sgRNA programmability and its tolerance to modifications.
  • Recent engineering advances have expanded the capabilities of sgRNAs in Cas9-mediated genome targeting.
  • Understanding sgRNA properties is crucial for optimizing CRISPR-based genome editing applications.

Conclusions:

  • The sgRNA is a critical determinant of the Cas9 platform's versatility and efficacy in genome targeting.
  • Continued engineering of sgRNAs holds significant potential for advancing CRISPR technology.
  • Future research should focus on further optimizing sgRNA design for precise and efficient genome editing.