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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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Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
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Related Experiment Video

Updated: Mar 1, 2026

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

Published on: May 25, 2018

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Chromatin accessibility and guide sequence secondary structure affect CRISPR-Cas9 gene editing efficiency.

Kristopher Torp Jensen1,2, Lasse Fløe1, Trine Skov Petersen1

  • 1Department of Biomedicine, Aarhus University, Denmark.

FEBS Letters
|June 6, 2017
PubMed
Summary

Gene editing efficiency with CRISPR-Cas9 is higher in accessible euchromatin than in inaccessible heterochromatin. Guide RNA secondary structure also impacts Cas9 editing activity, aiding tool design.

Keywords:
CRISPRgRNAchromatin accessibilityefficiency

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas9 systems are powerful genome editing tools, but variable on-target efficiencies hinder their application.
  • Understanding factors influencing CRISPR-Cas9 activity is crucial for improving gene editing precision.

Discussion:

  • Chromatin accessibility significantly impacts Cas9-mediated gene editing, with higher efficiency observed in euchromatin compared to heterochromatin.
  • This effect was validated across different cell types, including HEK293T, HeLa, and human fibroblasts.
  • Guide RNA secondary structure formation was identified as a key determinant of CRISPR-Cas9 editing efficiency.

Key Insights:

  • Gene editing is more efficient in open euchromatin regions than in condensed heterochromatin.
  • The secondary structure of guide RNA sequences directly influences the effectiveness of Cas9-mediated gene editing.
  • These findings provide critical insights into optimizing CRISPR-Cas9 system performance.

Outlook:

  • Further development of gRNA design tools can leverage the understanding of chromatin accessibility and guide sequence structure.
  • Improved gRNA design will enhance the reliability and efficiency of CRISPR-Cas9 genome editing applications.
  • This research paves the way for more predictable and effective gene editing strategies.