Potential limitations of the Sleeping Beauty transposon use in gene expression studies

Acta Biochim Pol. 2019 Jul 12;66(3):263-268. doi: 10.18388/abp.2019_2839.

Abstract

MCPIP2 is the least known member of the MCPIP family of proteins. Recently we have found that it is a new RNase involved in transcript turnover. However, the full spectrum of its cellular targets is still unidentified. To discover transcripts which are regulated by this protein we have employed Sleeping Beauty transposons. This tool allows for rapid generation of a stable transgenic cell line with inducible expression of the desired gene. In this study, we analysed how the Sleeping Beauty system itself influences expression of chosen genes, namely IL-6, Regnase-1 and VEGF. We found that the system alone may influence expression of IL-6. Our results indicate that Sleeping Beauty transposons should be used with caution in studies that are focused on changes in the transcript level.

MeSH terms

  • Aged
  • Brain Neoplasms / genetics
  • Brain Neoplasms / pathology
  • CRISPR-Cas Systems / genetics
  • Cell Line, Tumor
  • DNA Transposable Elements / genetics
  • Gene Editing
  • Gene Knockout Techniques
  • Glioblastoma / genetics
  • Glioblastoma / pathology
  • Humans
  • Interleukin-6 / genetics*
  • Male
  • Mutagenesis, Insertional / methods*
  • RNA, Messenger / genetics
  • Ribonucleases / genetics*
  • Transcription Factors / genetics*
  • Transcriptome / genetics*
  • Transfection
  • Vascular Endothelial Growth Factor A / genetics*

Substances

  • DNA Transposable Elements
  • IL6 protein, human
  • Interleukin-6
  • RNA, Messenger
  • Transcription Factors
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Ribonucleases
  • ZC3H12A protein, human
  • ZC3H12B protein, human