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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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Related Experiment Video

Updated: May 8, 2026

Mouse Genome Engineering Using Designer Nucleases
12:04

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Highly efficient targeted mutagenesis in mice using TALENs.

Sudeepta Kumar Panda1, Benedikt Wefers, Oskar Ortiz

  • 1Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany.

Genetics
|August 28, 2013
PubMed
Summary
This summary is machine-generated.

Researchers developed an efficient method for creating genetically modified mice using TALEN-95A mRNA and high-resolution melt analysis (HRMA). This accelerates the generation of mouse models for studying genetic diseases like frontotemporal lobar degeneration and amyotrophic lateral sclerosis.

Keywords:
C9ORF72FusTALENsdisease modelmouse mutantnucleaseone-cell embryo

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Area of Science:

  • Genetics
  • Molecular Biology
  • Genomics

Background:

  • Targeted mouse mutants are crucial for understanding gene function in health and disease.
  • Previous methods for generating mouse mutants were often time-consuming.
  • Transcription activator-like effector nucleases (TALENs) offer a powerful tool for genome engineering.

Purpose of the Study:

  • To establish a routine and efficient procedure for producing disease-related knockin and knockout mouse mutants.
  • To improve the TALEN system for faster and more reliable mutagenesis.
  • To validate a simple and universal genotyping tool for identifying TALEN-induced mutations.

Main Methods:

  • Utilized improved TALEN mRNAs with a plasmid-coded poly(A) tail (TALEN-95A) for enhanced mutagenesis.
  • Generated knockout mutants for the C9orf72 gene, a model for frontotemporal lobar degeneration.
  • Introduced patient-derived missense mutations into the Fused in Sarcoma (Fus) gene, relevant to amyotrophic lateral sclerosis, using TALENs and mutagenic oligodeoxynucleotides.
  • Validated high-resolution melt analysis (HRMA) for genotyping TALEN-induced mutants and assessing off-target effects.

Main Results:

  • TALEN-95A mutagenesis efficiently induced sequence deletions in 41% of pups for C9orf72 knockout.
  • Introduced Fus gene missense mutations at a rate of 6.8% using TALENs and mutagenic oligodeoxynucleotides.
  • HRMA proved to be a sensitive and universal tool for genotyping TALEN-induced mutants and their progeny.
  • HRMA analysis of off-target sites showed no evidence of undesired TALEN activity.

Conclusions:

  • The combination of TALEN-95A mRNAs and HRMA enables accelerated and routine production of mouse models.
  • This approach facilitates the study of genetic disease mechanisms by providing efficient generation of relevant mutants.
  • The validated methods enhance the speed and reliability of creating genetically engineered mouse models for research.