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Related Concept Videos

In-vitro Mutagenesis01:16

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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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Mice have long served as models for studying human biology and pathology because of their phylogenetic and physiological similarity with humans. They are also easy to maintain and breed in the laboratory, and hence, many inbred strains are now available for research. Studies on mice have contributed immeasurably to our understanding of cancer biology.
The development of transgenic, knockout, and knock-in mice has led to an exponential increase in their use as model organisms in research,...
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Related Experiment Video

Updated: Jul 10, 2025

Generation of Genetically Modified Mice through the Microinjection of Oocytes
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Genotyping Protocols for Genetically Engineered Mice.

Advait Limaye1, Kyoungin Cho2, Bradford Hall1

  • 1National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland.

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|November 20, 2023
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Summary
This summary is machine-generated.

This guide details methods for identifying genetically modified mouse models, focusing on polymerase chain reaction (PCR) techniques for both traditional and clustered regularly interspaced short palindromic repeat (CRISPR) gene editing. It provides essential protocols for accurate genetic trait verification in research settings.

Keywords:
CRISPR/Cas9genome editinggenotypingmouse models

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Area of Science:

  • * Genetics and Genomics
  • * Mammalian Model Systems
  • * Molecular Biology

Background:

  • * Laboratory mice are crucial for studying gene function and human disease modeling.
  • * Advancements in gene editing technologies like CRISPR have enabled precise mouse genome manipulation.
  • * Standardized methods are essential for consistent identification of genetically modified mouse models across research facilities.

Purpose of the Study:

  • * To provide comprehensive guidelines for identifying genetically modified mouse models.
  • * To detail endpoint polymerase chain reaction (PCR) methods for genotyping.
  • * To outline strategies for identifying models created using newer gene-editing technologies, including CRISPR.

Main Methods:

  • * Genomic DNA extraction from various mouse tissues and samples (tail biopsy, ear punch, blastocysts, semen, blood, buccal swabs).
  • * Endpoint polymerase chain reaction (PCR) for routine genotyping.
  • * CRISPR-specific assays including T7E1/Surveyor, off-target mutation detection, and gene knock-in/deletion analysis.

Main Results:

  • * Established protocols for reliable DNA purification from diverse biological samples.
  • * Demonstrated the efficacy of endpoint PCR for routine genotyping of genetically modified mice.
  • * Presented methods for verifying CRISPR-mediated edits, including insertion/deletion detection and off-target analysis.

Conclusions:

  • * Standardized molecular techniques are vital for accurate characterization of genetically modified mouse models.
  • * Endpoint PCR remains a robust method for genotyping, complemented by specialized assays for CRISPR-edited models.
  • * These guidelines facilitate consistent and reproducible research using genetically engineered mice.