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Related Concept Videos

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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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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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Related Experiment Video

Updated: Jun 23, 2025

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis
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CRISPR/Pepper-tDeg: A Live Imaging System Enables Non-Repetitive Genomic Locus Analysis with One Single-Guide RNA.

Meng Chen1, Xing Huang1, Yakun Shi1

  • 1Guangdong Provincial Key Laboratory of Sensing Technology and Biomedical Instrument, School of Biomedical Engineering, Shenzhen Campus of Sun Yat-Sen University, Sun Yat-Sen University, Shenzhen, 518107, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|June 26, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed CRISPR/Pepper-tDeg, an advanced genomic imaging system. This system significantly enhances signal-to-noise ratio for visualizing repetitive and non-repetitive DNA sequences in living cells.

Keywords:
CRISPR‐Cas9fluorogenic proteingenomic loci labelinglive cell imagingnon‐repetitive sequences

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • CRISPR-based genomic imaging enables spatiotemporal visualization of DNA in living cells.
  • Existing systems face challenges with low signal-to-noise ratio (SNR), especially for non-repetitive genomic loci.
  • Improved imaging techniques are crucial for understanding genome organization and dynamics.

Purpose of the Study:

  • To develop an efficient CRISPR-based genomic imaging system with enhanced SNR.
  • To enable sensitive imaging of repetitive and non-repetitive genomic loci.
  • To reduce technical barriers for studying genome organization and dynamics.

Main Methods:

  • Engineered CRISPR sgRNA scaffolds with degron-binding Pepper aptamers.
  • Utilized fluorogenic proteins fused with Tat peptide derived degron domain (tDeg).
  • Implemented target-dependent stability switches for sgRNA and fluorogenic proteins.

Main Results:

  • CRISPR/Pepper-tDeg achieved a 5-fold higher SNR for telomere imaging compared to CRISPR/MS2-MCP.
  • Successfully demonstrated simultaneous labeling and tracking of telomeres and centromeres.
  • Extended the system for non-repetitive sequence imaging with improved SNR using split fluorescent proteins.

Conclusions:

  • CRISPR/Pepper-tDeg offers sensitive and efficient genomic imaging with high SNR.
  • The system simplifies sgRNA design and plasmid construction, lowering technical barriers.
  • Shows significant potential for biological research, clinical diagnosis, and therapy.