A Rapid and Cost-Effective Method for Genotyping Genome-Edited Animals: A Heteroduplex Mobility Assay Using Microfluidic Capillary Electrophoresis

Vanessa Chenouard^{a, b, #},
Lucas Brusselle^{a, b, #},
Jean-Marie Heslan^{a, c},
Séverine Remy^{a, b},
Séverine Ménoret^{a, b},
Claire Usal^{a, b},
Laure-Hélène Ouisse^{a, b},
Tuan Huy NGuyen^{a, c},
Ignacio Anegon^{a, b},
Laurent Tesson^{a, b
, ,}

a.
Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France
b.
Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France
c.
GenoCellEdit, INSERM UMR 1064-ITUN, CHU Nantes, Nantes 44093, France

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Corresponding author: E-mail address: laurent.tesson@univ-nantes.fr (Laurent Tesson)

These two authors contributed equally to this work.

摘要

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Abstract: The recent emergence and application of engineered endonucleases have led to the development of genome editing tools capable of rapidly implementing various targeted genome editions in a wide range of species. Moreover, these novel tools have become easier to use and have resulted in a great increase of applications. Whilst gene knockout (KO) or knockin (KI) animal models are relatively easy to achieve, there is a bottleneck in the detection and analysis of these mutations. Although several methods exist to detect these targeted mutations, we developed a heteroduplex mobility assay on an automated microfluidic capillary electrophoresis system named HMA-CE in order to accelerate the genotyping process. The HMA-CE method uses a simple PCR amplification of genomic DNA (gDNA) followed by an automated capillary electrophoresis step which reveals a heteroduplexes (HD) signature for each mutation. This allows efficient discrimination of wild-type and genome-edited animals down to the single base pair level.
- Genome editing /
- Genotyping /
- Heteroduplex mobility assay /
- Capillary electrophoresis
These two authors contributed equally to this work.
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doi: 10.1016/j.jgg.2016.04.005

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A Rapid and Cost-Effective Method for Genotyping Genome-Edited Animals: A Heteroduplex Mobility Assay Using Microfluidic Capillary Electrophoresis

Corresponding author: E-mail address: laurent.tesson@univ-nantes.fr (Laurent Tesson)

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doi: 10.1016/j.jgg.2016.04.005

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A Rapid and Cost-Effective Method for Genotyping Genome-Edited Animals: A Heteroduplex Mobility Assay Using Microfluidic Capillary Electrophoresis

Corresponding author: E-mail address: laurent.tesson@univ-nantes.fr (Laurent Tesson)

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