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

J Genet Genomics. 2016 May 20;43(5):341-8. doi: 10.1016/j.jgg.2016.04.005. Epub 2016 May 4.


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.

Keywords: Capillary electrophoresis; Genome editing; Genotyping; Heteroduplex mobility assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cost-Benefit Analysis*
  • Electrophoresis, Capillary / economics
  • Electrophoresis, Capillary / instrumentation*
  • Gene Editing*
  • Genotyping Techniques / economics*
  • Genotyping Techniques / instrumentation
  • Lab-On-A-Chip Devices*
  • Mutation
  • Rats
  • Time Factors