Design and characterization of a salicylic acid-inducible gene expression system for Jurkat cells

J Biotechnol. 2022 Feb 20:346:11-14. doi: 10.1016/j.jbiotec.2022.01.003. Epub 2022 Jan 18.

Abstract

With continued progress in cell and gene therapies, there is an immediate need for exogenously tunable gene expression systems with safe and predictable behavior in specific human cell types. Here, we demonstrate the ability of the salicylic acid (SA)-inducible MarR repressor protein from Escherichia coli to regulate target gene expression in a human T lymphocyte cell line. Two lentiviral vectors, one encoding an enhanced green fluorescent protein (EGFP) reporter cassette and the other a repressor cassette, were sequentially transduced into Jurkat cells, using fluorescence-activated cell sorting (FACS) to isolate stable Jurkat progeny. As a result, EGFP expression was repressed by MarR and was inducible upon the addition of SA (~1.3 fold). This represents the first example of functional expression of bacterial MarR in mammalian cells, and opens the possibility for further development of regulated, SA-tunable gene expression system for T-cells.

Keywords: FACS; Gene repression; Inducible promoter; Jurkat cells; Lentiviral transduction; Transcriptional factors.

MeSH terms

  • Animals
  • Gene Expression
  • Genetic Vectors* / genetics
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Jurkat Cells
  • Lentivirus* / genetics
  • Salicylic Acid

Substances

  • Green Fluorescent Proteins
  • Salicylic Acid