Exposure of cells to ionising radiation results in the activation of specif
ic transcriptional control (CArG) elements within the early growth response
I (Egr1) gene promoter leading to increased gene expression. As part of a
study investigating the potential use of these elements in radiation-contro
lled gene therapy vectors, we have incorporated their sequences into a synt
hetic gene promoter and assayed for the ability to induce expression of a d
ownstream reporter gene following irradiation. In vector-transfected MCF-7
breast adenocarcinoma cells, the synthetic promoter was more effective than
the wild-type Egr1 counterpart in up-regulating expression of the reporter
gene after exposure to a single 5 Gy dose, and equally effective as the wi
ld-type in U87-MG glioma cells. The level of gene expression achieved using
the synthetic promoter was dependent on the inducing radiation dose for bo
th U87-MG and MCF-7 cells, being maximal at 3 Gy and decreasing at 5 and 10
Gy. Furthermore, induction could be repeated by additional radiation treat
ments. The latter indicates that up-regulation should be additive during fr
actionated radiotherapy schedules. To demonstrate the potential clinical be
nefit of such an approach, the synthetic promoters were also shown to drive
expression of the herpes simplex virus thymidine kinase gene, leading to e
nhanced cell killing in the presence of the prodrug ganciclovir (GCV) when
compared with cells treated with radiation alone. Our results demonstrate t
hat the synthetic promoter is responsive to low doses of ionising radiation
and therefore isolated CArG elements function as radiation-mediated transc
riptional enhancers outside their normal sequence context. The continued de
velopment and optimisation of such radiation-responsive synthetic promoters
is expected to make a valuable contribution to the development of future r
adiation-responsive vectors for cancer gene therapy.