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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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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 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.
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Related Experiment Video

Updated: Sep 29, 2025

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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Boosting targeted genome editing using the hei-tag.

Thomas Thumberger1, Tinatini Tavhelidse-Suck1,2, Jose Arturo Gutierrez-Triana1

  • 1Centre for Organismal Studies (COS), Heidelberg University, Heidelberg, Germany.

Elife
|March 25, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed the "hei-tag" to significantly improve CRISPR/Cas9 genome editing efficiency. This novel tag enhances gene editing accuracy and speed when delivered via mRNA, benefiting various model systems.

Keywords:
CRISPRCasOryzias latipesbase editinggeneticsgenomicsmousenuclear localizationoryzias latipestargeted genome editingzebrafish

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 is a powerful genome editing tool crucial for research and translation.
  • Current CRISPR/Cas9 systems require optimization for nuclear delivery and detection.
  • Enhancing editing efficiency and immediate activity remains a key challenge.

Purpose of the Study:

  • To introduce and evaluate the 'hei-tag' for boosting CRISPR/Cas genome editing efficiency.
  • To assess the impact of the hei-tag on targeting accuracy and editing outcomes.
  • To demonstrate the broad applicability of the hei-tag across different model systems.

Main Methods:

  • Development of the 'hei-tag' (high efficiency tag) by fusing a myc-tag with an optimized nuclear localization signal (NLS).
  • Fusion of the hei-tag to Cas9 and C-to-T base editors.
  • Delivery of mRNA encoding hei-tag fused proteins for genome editing experiments.
  • Testing in various model systems including fish, mammals, and cell cultures.

Main Results:

  • The hei-tag significantly enhanced CRISPR/Cas9 and base editor targeting efficiency.
  • Increased bi-allelic editing rates and reduced allele variance were observed.
  • The tag demonstrated immediate activity, even at early developmental stages.
  • Hei-tag efficacy was confirmed across diverse model organisms and cell culture.

Conclusions:

  • The hei-tag is a simple yet powerful addition to enhance CRISPR/Cas genome editing tools.
  • It improves editing efficiency and accuracy when delivered as mRNA.
  • The hei-tag offers a versatile solution for upgrading existing systems and developing new, highly efficient genome editing applications.