TY - JOUR
T1 - Design and characterization of a salicylic acid-inducible gene expression system for Jurkat cells
AU - Doshi, Aarti
AU - Bandey, Irfan
AU - Nevozhay, Dmitry
AU - Varadarajan, Navin
AU - Cirino, Patrick C.
N1 - Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/2/20
Y1 - 2022/2/20
N2 - 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.
AB - 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.
KW - FACS
KW - Gene repression
KW - Inducible promoter
KW - Jurkat cells
KW - Lentiviral transduction
KW - Transcriptional factors
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U2 - 10.1016/j.jbiotec.2022.01.003
DO - 10.1016/j.jbiotec.2022.01.003
M3 - Article
C2 - 35051448
AN - SCOPUS:85123711865
SN - 0168-1656
VL - 346
SP - 11
EP - 14
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -