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Related Experiment Video

Updated: Aug 4, 2025

Mouse Genome Engineering Using Designer Nucleases

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Genotyping Genome-Edited Founders and Subsequent Generation.

Matthew Mackenzie¹, Alex Fower¹, Alasdair J Allan¹

¹The Mary Lyon Centre, MRC Harwell, Didcot, Oxon, UK.

Methods in Molecular Biology (Clifton, N.J.)

|March 30, 2023

Summary

Developing precise molecular assays and genotyping strategies is crucial for identifying and validating genetically engineered founder animals produced using targeted nucleases in early embryos.

Keywords:

Allele QC CRISPR-Cas9 Embryo/zygote engineering Genetic engineering Genome editing Genotyping Mouse models Transgenesis

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Published on: April 11, 2019

Area of Science:

Molecular biology
Genetics
Developmental biology

Background:

Targeted nucleases enable direct genetic mutation induction in early embryos.
However, resulting DNA repair is unpredictable, leading to mosaic founder animals.

Purpose of the Study:

To present molecular assays and genotyping strategies.
To support screening of first-generation founder animals.
To validate positive animals in subsequent generations based on mutation type.

Main Methods:

Development of molecular assays for mutation detection.
Establishment of genotyping strategies for founder screening.
Application of methods for subsequent generation validation.

Main Results:

The presented assays and strategies facilitate the identification of potential founder animals.
These methods allow for the validation of specific mutations in subsequent generations.
The approach addresses the challenge of mosaicism in genetically engineered animals.

Conclusions:

Robust molecular assays and genotyping are essential for efficient genetic engineering workflows.
These strategies enable reliable characterization of mutations in founder animals and their offspring.
The findings support the advancement of genetic engineering applications in developmental biology.