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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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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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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator
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Clonal analysis by tunable CRISPR-mediated excision.

Anna F Gilles1,2,3, Johannes B Schinko1,3,4, Magdalena I Schacht5,6

  • 1Institut de Génomique Fonctionnelle de Lyon (IGFL), École Normale Supérieure de Lyon, 32 avenue Tony Garnier, 69007 Lyon, France anna.gilles@trigenes.com johannes.schinko@trigenes.com michalis.averof@ens-lyon.fr.

Development (Cambridge, England)
|December 16, 2018
PubMed
Summary
This summary is machine-generated.

Valcyrie uses CRISPR technology for precise cell marking in model organisms, offering tunable clone induction frequencies for genetic analysis. This method enhances mosaic genetic analysis by controlling cell marking efficiency.

Keywords:
CRISPRClonal analysisGenetic mosaicsInsectTribolium

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Area of Science:

  • Developmental Biology
  • Genetics
  • Molecular Biology

Background:

  • Cre/lox and Flp/FRT systems are standard for clonal marking and mosaic genetic analysis in model organisms.
  • Controlling clone induction frequency is crucial but challenging with existing methods.
  • Need for adaptable tools to study cell fate, morphology, and growth properties.

Purpose of the Study:

  • Introduce Valcyrie, a novel CRISPR-mediated method for clonal marking.
  • Enable tunable control over clone induction frequency for precise genetic analysis.
  • Adapt clonal analysis techniques for established and emerging model organisms.

Main Methods:

  • Replaced conventional recombinase-mediated cassette excision with CRISPR-mediated excision.
  • Utilized tunable CRISPR efficiency by manipulating guide RNA sequence complementarity.
  • Established and validated the Valcyrie method in the beetle model organism, *Tribolium castaneum*.

Main Results:

  • Demonstrated successful clonal marking using CRISPR-mediated excision.
  • Showcased predictable tuning of clone marking frequency.
  • Generated embryos with single marked clones, enabling detailed analysis.

Conclusions:

  • Valcyrie offers a powerful and tunable alternative to traditional clonal marking systems.
  • The method facilitates precise mosaic genetic analysis by controlling induction frequency.
  • Valcyrie is broadly applicable to various experimental settings and model systems.