MosSCI-mediated exogenous gene expression is modulated by genomic positioning

Biotechnol J. 2023 Sep;18(9):e2300062. doi: 10.1002/biot.202300062. Epub 2023 May 22.

Abstract

Although the Mos1-mediated single-copy insertion (MosSCI) technique has been widely used to generate stable transgenic Caenorhabditis elegans strains, the link between stability of expression and integration site still needs to be explored. Here, experimental evidence is provided that transgenes are not able to match the level of transcription of their native counterpart, and that insertions at certain locations can result in an external stress-mediated increase in expression. Insertion site ttTi5605 on chromosome II was shown to be a superior location, at least when introducing reproduction related genes. Thus, this study provides a reference for the selection of an optimal site for MosSCI which provides acceptable expression performance whilst minimizing undesirable secondary effects.

Keywords: Caenorhabditis elegans; genetic engineering; single-copy insertion; transcription.

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Caenorhabditis elegans* / genetics
  • Caenorhabditis elegans* / metabolism
  • Gene Expression
  • Genome*
  • Genomics
  • Mutagenesis, Insertional
  • Transgenes / genetics