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Related Concept Videos

Homologous Recombination02:31

Homologous Recombination

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...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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...
Mismatch Repair01:36

Mismatch Repair

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Updated: May 29, 2026

Mosaic Zebrafish Transgenesis for Functional Genomic Analysis of Candidate Cooperative Genes in Tumor Pathogenesis
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Published on: March 31, 2015

Tuning mitotic recombination with patterned DNA nicks for precision mosaic analysis.

Yifan Shen1,2, Ann T Yeung1,2, Bei Wang1,2

  • 1Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853.

Proceedings of the National Academy of Sciences of the United States of America
|May 27, 2026
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 gene editing can cause DNA damage. Researchers developed safer CRISPR nickases for Drosophila mosaic analysis, using single-strand nicks to induce genetic recombination with reduced toxicity and improved reliability.

Keywords:
CRISPRDrosophilaMAGICmosaic analysisnickase

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Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Background:

  • CRISPR/Cas9 gene editing is vital for in vivo genetics.
  • DNA double-strand breaks from CRISPR/Cas9 cause cytotoxicity and mutagenesis, limiting its use.
  • Safer alternatives are needed for precise genetic analysis.

Purpose of the Study:

  • To establish CRISPR-derived nickases as safer alternatives for inducing mitotic recombination in Drosophila.
  • To investigate the parameters controlling nick-induced recombination.
  • To develop a toolkit for tissue-specific nickase generation.

Main Methods:

  • Utilized Cas9-derived nickases to induce single-strand DNA breaks in Drosophila.
  • Systematically analyzed the effect of gRNA nicking patterns on recombination frequency.
  • Developed a mechanistic model for nick-induced crossover.

Main Results:

  • Single-strand nicks are sufficient to generate mosaic clones.
  • Recombination frequency is controllable via gRNA nicking patterns.
  • Two distant nicks on the same DNA strand synergistically increased recombination over ninefold compared to a single nick.

Conclusions:

  • Nickase-based mosaic analysis using gRNA-induced crossing-over is a superior method for high-fidelity clonal analysis.
  • This approach enables more precise investigation of gene function in development and disease.
  • Established a versatile toolkit for generating tissue-specific nickases.