Extremely high and specific activity of DNA enzymes in cells with a Philadelphia chromosome.
BACKGROUND
Chronic myelogenous leukemia (CML) results from chromosome 22 translocations (the Philadelphia chromosome) that creates BCR-ABL fusion genes, which encode two abnormal mRNAs (b3a2 and b2a2). Various attempts to design antisense oligonucleotides that specifically cleave abnormal L6 BCR-ABL fusion mRNA have not been successful. Because b2a2 mRNA cannot be effectively cleaved by hammerhead ribozymes near the BCR-ABL junction, it has proved very difficult to engineer specific cleavage of this chimeric mRNA. Nonspecific effects associated with using antisense molecules make the use of such antisense molecules questionable.


RESULTS
The usefulness of DNA enzymes in specifically suppressing expression of L6 BCR-ABL mRNA in mammalian cells is demonstrated. Although the efficacy of DNA enzymes with natural linkages decreased 12 hours after transfection, partially modified DNA enzymes, with either phosphorothioate or 2'-O-methyl groups at both their 5' and 3' ends, remained active for much longer times in mammalian cells. Moreover, the DNA enzyme with only 2'-O-methyl modifications was also highly specific for abnormal mRNA.


CONCLUSIONS
DNA enzymes with 2'-O-methyl modifications are potentially useful as gene-inactivating agents in the treatment of diseases such as CML. In contrast to conventional antisense DNAs, some of the DNA enzymes used in this study were highly specific and cleaved only abnormal BCR-ABL mRNA.
Warashina
M
M
National Institute for Advanced Interdisciplinary Research, AIST, Tsukuba Science City 305-8562, Japan.
Kuwabara
T
T
Nakamatsu
Y
Y
Taira
K
K
D002478:Cells, Cultured; D004795:Enzyme Stability; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D010677:Philadelphia Chromosome; D012333:RNA, Messenger / Q000378:metabolism; D013379:Substrate Specificity
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl