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

Aarti Doshi, Irfan Bandey, Dmitry Nevozhay, Navin Varadarajan, Patrick C. Cirino

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)11-14
Number of pages4
JournalJournal of Biotechnology
Volume346
DOIs
StatePublished - Feb 20 2022

Keywords

  • FACS
  • Gene repression
  • Inducible promoter
  • Jurkat cells
  • Lentiviral transduction
  • Transcriptional factors

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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