Construction of a ribozyme directed against human interleukin-6 mRNA:
evaluation of its catalytic activity in vitro and in vivo
M Mahieu, R Deschuyteneer, D Forget, P Vandenbussche and J Content
Institut Pasteur du Brabant, Department of Virology, Brussels, Belgium.
We have designed a ribozyme (Rz) that cleaves human interleukin-6 (IL- 6)
mRNA in vivo. This Rz was tested in vitro, and was found to give expected
size fragments. It was then incorporated into a mammalian expression vector
containing the constitutive cytomegalovirus (CMV) immediate early promoter
and transfected into human U amniotic cells (UAC). Cell clones that stably
express this catalytic RNA have been obtained. Some of them displayed a
marked reduction of tumor necrosis factor (TNF)-induced IL-6 production.
Their reduced ability to express IL-6 was related to the amount of Rz they
produced and to the extent of IL-6 mRNA cleavage as observed by a
ribonuclease protection assay. These data provide a method to study further
the role of IL-6 production in various biologic situations, and suggest the
feasibility of developing Rzs directed against various cytokines to study
their biologic role and mechanism of action.
Volume 84,
Issue 11,
pp. 3758-3765,
12/01/1994
Copyright © 1994 by The American Society of Hematology
