A deterministic genotyping workflow reduces waste of transgenic individuals by two-thirds

Sci Rep. 2021 Jul 28;11(1):15325. doi: 10.1038/s41598-021-94288-0.

Abstract

We present a deterministic workflow for genotyping single and double transgenic individuals directly upon nascence that prevents overproduction and reduces wasted animals by two-thirds. In our vector concepts, transgenes are accompanied by two of four clearly distinguishable transformation markers that are embedded in interweaved, but incompatible Lox site pairs. Following Cre-mediated recombination, the genotypes of single and double transgenic individuals were successfully identified by specific marker combinations in 461 scorings.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Embryo, Nonmammalian
  • Female
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Gene Expression
  • Genetic Engineering / economics
  • Genetic Engineering / methods*
  • Genetic Markers
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism*
  • Genotyping Techniques*
  • Heterozygote
  • Histones / genetics
  • Histones / metabolism
  • Homozygote
  • Integrases / genetics*
  • Integrases / metabolism
  • Male
  • Microscopy, Fluorescence
  • Promoter Regions, Genetic
  • Tribolium / genetics*
  • Tubulin / genetics
  • Tubulin / metabolism

Substances

  • Fluorescent Dyes
  • Genetic Markers
  • Histones
  • Tubulin
  • Cre recombinase
  • Integrases