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  • Title: Specificity of ribozyme designed for mutated DHFR mRNA.
    Author: Kobayashi H, Kim N, Halatsch ME, Ohnuma T.
    Journal: Biochem Pharmacol; 1994 Apr 29; 47(9):1607-13. PubMed ID: 8185675.
    Abstract:
    When MOLT-3 human acute leukemia cells were exposed sequentially to trimetrexate (TMQ) and then to methotrexate (MTX), the cells became resistant to antifolate. We designated this subline MOLT-3/TMQ800-MTX10,000. This cell line was found to contain two point mutations in the dihydrofolate reductase (DHFR) gene: a T-->C transition at nucleotide 95 in codon 31, and a T-->A transition at nucleotide 100 in codon 33. In an attempt to specifically inhibit these double-mutated cells, we synthesized a ribozyme which perfectly base-paired with the double-mutated DHFR mRNA. We found that the ribozyme for the double-mutated DHFR mRNA not only cleaved the mutated DHFR RNA, but also efficiently cleaved the wild-type RNA substrate. This observation suggests proceeding with caution when using a ribozyme against a mutated mRNA of an essential enzyme as a specific means of treatment.
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