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Ribozyme-mediated modulation of human O6-methylguanine-DNA methyltransferase expression
P M Potter1, L C Harris, J S Remack
1Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Abstract:
A synthetic oligonucleotide containing ribozyme sequences targeted to the 5' region of the human O6-methylguanine-DNA methyltransferase (MGMT) mRNA has been constructed. This ribozyme demonstrates cleavage activity in vitro in the presence of Mg2+. To determine whether this ribozyme can function in vivo, HeLa CCL2 cells were transfected with a mammalian expression vector containing the ribozyme sequence. Following selection and expansion of individual transfectants, a stable clone was isolated that lacks both MGMT mRNA and protein. Molecular analysis of this cell line indicates that in vivo cleavage of MGMT mRNA is responsible for the lack of MGMT activity.