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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice
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Floxin, a resource for genetically engineering mouse ESCs.

Veena Singla1, Julie Hunkapiller, Nicole Santos

  • 1Department of Biochemistry and Biophysics, Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California, USA.

Nature Methods
|December 8, 2009
PubMed
Summary
This summary is machine-generated.

The Floxin method allows for efficient and precise gene editing in mouse embryonic stem cells (ESCs). This technology enables high-throughput modification of numerous genes using existing gene trap cell lines.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Stem Cell Research

Background:

  • Gene trap technology is crucial for understanding gene function.
  • Existing gene trap mouse embryonic stem cell (ESC) lines offer a valuable resource.
  • Targeted modification of these loci is essential for advanced functional genomics.

Purpose of the Study:

  • To develop a method for efficient and precise modification of gene trap loci in mouse ESCs.
  • To enable high-throughput genetic manipulation using existing compatible gene trap cell lines.

Main Methods:

  • Utilized the Floxin method involving Cre recombinase and a shuttle vector (pFloxin).
  • Demonstrated reversion of gene trap mutations and insertion of new DNA sequences.
  • Applied the method to a large collection of compatible gene trap ESC lines.

Main Results:

  • Achieved highly efficient and precise targeted modification of gene trap loci.
  • Successfully reverted existing mutations and inserted novel DNA sequences.
  • Validated the applicability of the Floxin method to over 24,000 gene trap cell lines.

Conclusions:

  • The Floxin method provides a powerful tool for genetic manipulation in mouse ESCs.
  • This technology facilitates high-throughput functional genomics studies.
  • Enables rapid and precise characterization of gene function through targeted DNA modification.