title: A Rapid and Cost-Effective Method for Genotyping Genome-Edited Animals: A Heteroduplex Mobility Assay Using Microfluidic Capillary Electrophoresis.
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.
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Chenouard
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Lucas
authors.initials: L
authors.lastname: Brusselle
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; GenoCellEdit, INSERM UMR 1064-ITUN, CHU Nantes, Nantes 44093, France.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Heslan
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Remy
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Ménoret
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Usal
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Laure-Hélène
authors.initials: LH
authors.lastname: Ouisse
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; GenoCellEdit, INSERM UMR 1064-ITUN, CHU Nantes, Nantes 44093, France.
authors.forename: Tuan Huy
authors.initials: TH
authors.lastname: NGuyen
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Anegon
authors.affiliation: Institut National de la Santé Et de la Recherche Médicale UMR 1064-Institut Transplantation Urologie-Néphrologie, Centre Hospitalier Universitaire de Nantes, Nantes 44093, France; Transgenesis Rat ImmunoPhenomic Platform, Structure Fédérative de Recherche François Bonamy, Centre National de Recherche Scientifique UMS3556, Nantes 44093, France. Electronic address: laurent.tesson@univ-nantes.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Tesson
mesh_terms: D000818:Animals; D003362:Cost-Benefit Analysis; D019075:Electrophoresis, Capillary / Q000191:economics / Q000295:instrumentation*; D000072669:Gene Editing; D060005:Genotyping Techniques / Q000191:economics* / Q000295:instrumentation; D056656:Lab-On-A-Chip Devices; D009154:Mutation; D051381:Rats; D013997:Time Factors
keywords: Capillary electrophoresis; Genome editing; Genotyping; Heteroduplex mobility assay