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Related Concept Videos

phiC31-Integrase-Mediated Site-Directed Transgene Integration: A Microinjection Technique for Site-Specific Transgene Integration into an Anopheles Vector04:21

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The functional genomic toolkit for the parasitic nematodes Strongyloides stercoralis and Strongyloides ratti includes transgenesis, CRISPR/Cas9-mediated mutagenesis, and RNAi. This protocol will demonstrate how to use intragonadal microinjection to introduce transgenes and CRISPR components into S. stercoralis and...
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Stably transgenic Hydra are made by microinjection of plasmid DNA into embryos followed by random genomic integration and asexual propagation to establish a uniform line. Transgenic Hydra are used to track cell movements, overexpress genes, study promoter function, or knock down gene expression using RNAi.
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Related Experiment Video

Updated: Jan 19, 2026

phiC31-Integrase-Mediated Site-Directed Transgene Integration: A Microinjection Technique for Site-Specific Transgene Integration into an Anopheles Vector
04:21

phiC31-Integrase-Mediated Site-Directed Transgene Integration: A Microinjection Technique for Site-Specific Transgene Integration into an Anopheles Vector

2.8K

Integrase-Mediated Targeted Transgenics Through Pronuclear Microinjection.

Ruby Yanru Chen-Tsai1

  • 1Applied StemCell, Inc., Milpitas, CA, USA. Ruby.tsai@appliedstemcell.com.

Methods in Molecular Biology (Clifton, N.J.)
|September 13, 2019
PubMed
Summary

A new TARGATT™ system enables site-specific transgenic mice creation via microinjection. This method efficiently inserts single-copy transgenes into targeted loci, facilitating rapid gene function and regulation studies.

Keywords:
H11 locusPhiC31 integrasePronuclear microinjectionRosa26 knockinSite-specific transgenesisTARGATT™

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Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Animal Models

Background:

  • Transgenic technology is crucial for disease mechanism research.
  • Traditional microinjection methods lead to random, multi-copy insertions, causing position effects and variable expression.
  • Homologous recombination in embryonic stem cells offers targeted insertion but is labor-intensive.

Purpose of the Study:

  • To introduce an efficient, site-specific method for generating transgenic mice.
  • To overcome limitations of random insertion and laborious targeted integration techniques.
  • To facilitate precise dissection of gene function and regulation in vivo.

Main Methods:

  • Development and application of the TARGATT™ (target attP) integrase-based system.
  • Site-specific insertion of single-copy transgenes into predetermined chromosomal loci via pronuclear microinjection.
  • Utilizing various promoters for high-level, tissue-specific, or inducible transgene expression.

Main Results:

  • Achieved high efficiency (up to 40%) of site-specific transgene insertion.
  • Ensured faithful transmission of transgenes through generations.
  • Enabled rapid generation of site-specific transgenic mice (as fast as 3 months).

Conclusions:

  • The TARGATT™ system significantly advances murine transgenesis.
  • Enables precise structure/function dissection of mammalian gene function and regulation.
  • Offers a faster and more reliable alternative to existing transgenic mouse generation methods.