A Method for Screening Antisense Oligodeoxyribonucleotides Effective for mRNA Translation-Arrest.

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概要

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• A transcription and translation coupled reticulocyte lysate system was established for rapid screening of antisense oligodeoxyribonucleotides (ODNs) to determine which are most effective for mRNA translation-arrest. A plasmid containing the target cDNA under the control of the T7 (or SP6) promoter was added to the lysate system in the presence of the T7 (or SP6) RNA polymerase, RNase H, and the antisense ODN under test. Transcription and translation were accomplished in a one-tube reaction. Translation-arrest caused by antisense ODN was evaluated in terms of the amounts of de novo-synthesized, [35S]-methionine or [35S] cysteine labeled target protein measured by gel electrophoresis and autoradiography. The properties of this system and optimal reaction conditions for use in antisense ODN screening were determined. Our method is simpler and more rapid than other in vitro screening methods.

• 社団法人 日本生化学会の論文

著者

• Au Lo-chun

  Graduate Institute Of Medical Biotechnology National Yang-ming University

• Lin Shwu-bin

  Graduate Institute Of Medical Technology National Taiwan University Hospital

• Wu Jaw-ching

  Institute Of Clinical Medicine School Of Medicine National Yang-ming University

• CHOO Kong-Bung

  Graduate Institute of Medical Biotechnology National Yang-Ming University

• Chen Trong-zong

  Institute Of Clinical Medicine School Of Medicine National Yang-ming University

• LIN Shwu-Bin

  Graduate Institute of Medical Technology, College of Medicine, National Taiwan University

関連論文

• Antisense Oligodeoxynucleotides of IGF-II Selectively Inhibit Growth of Human Hepatoma Cells Overproducing IGF-II
• A Method for Screening Antisense Oligodeoxyribonucleotides Effective for mRNA Translation-Arrest^1
• Inhibition of α-Fetoprotein Production in a Hepatoma Cell Line by Antisense Oligonucleotide Analogues^1
• A Method for Screening Antisense Oligodeoxyribonucleotides Effective for mRNA Translation-Arrest.
• Antisense Oligodeoxynucleotides of IGF-II Selectively Inhibit Growth of Human Hepatoma Cells Overproducing IGF-II.

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