Application of the colorimetric loop-mediated isothermal amplification (LAMP) technique for genotyping Cre-driver mice

J Vet Med Sci. 2022 Apr 13;84(4):507-510. doi: 10.1292/jvms.21-0658. Epub 2022 Feb 27.

Abstract

The Cre-loxP system is widely used to investigate the cell-type specific roles of genes of interest. Cre-driver mice are required for cell-type specific knockout during the Cre-loxP reaction. To maintain Cre-driver mouse strains, Polymerase chain reaction (PCR)-oriented genotyping targeting the Cre gene cassette is usually conducted. In this study, we instead applied a colorimetric loop-mediated isothermal amplification (LAMP) method for Cre-genotyping. Among four sets of primers designed by the in silico program, one set effectively amplified the Cre cassette of three Cre-driver strains, but not of C57BL/6 mice. This LAMP-oriented method reduces assay time by less than half compared to the PCR-based method, and can be carried out using a conventional isothermal incubator. Applying this LAMP method may accelerate genotyping of Cre-driver mice.

Keywords: Cre; conditional knockout; genotyping; loop-mediated isothermal amplification.

MeSH terms

  • Animals
  • Colorimetry* / veterinary
  • Genotype
  • Integrases
  • Mice
  • Mice, Inbred C57BL
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques* / methods
  • Nucleic Acid Amplification Techniques* / veterinary
  • Sensitivity and Specificity

Substances

  • Cre recombinase
  • Integrases

Supplementary concepts

  • LAMP assay