Design and Construction of bcr3/abl2 Specific Ribozyme and Further Study in Specifically Inhibiting Growth and Colony Formation of K562 Cells

LI Man wen,HE Fu chu

Chinese Journal of Biochemistry and Molecular Biology ›› 2000, Vol. 16 ›› Issue (01) : 10-16.

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PDF(275 KB)
Chinese Journal of Biochemistry and Molecular Biology ›› 2000, Vol. 16 ›› Issue (01) : 10-16.
Article

Design and Construction of bcr3/abl2 Specific Ribozyme and Further Study in Specifically Inhibiting Growth and Colony Formation of K562 Cells

  • LI Man wen,HE Fu chu
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Abstract

With the aid of computer software,two hammerhead ribozymes (RZ0&RZT) were designed,which targeted to the UUC site on fusion junction of bcr3/abl2 fusion gene according to the promoted ribozyme structure model published by Haseloff and Gerlach.The cleavage substrate of RZ0&RZT came from a fraction of bcr3/abl2 fusion gene within which the target site was included.The in vitro cleavage activity of ribozyme RZ0&RZT was assessed. Further,the RZT gene was cloned into pCEP4, which could transiently express in eukaryotic cells,then transferred into K562 cells. Results showed that those ribozymes originated from expression vector RZT/pCEP4 and the in vitro transcripts were capable of specifically inhibiting the growth and colony formaction and initiating apoptosis of K562 cells. Finally,it was observed that the inhibition activity of ribozyme RZT was higher than that of both ribozyme RZ0 and relevant ASON.

Key words

Hammerhead ribozyme / bcr3/abl2 / Apoptosis

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LI Man wen,HE Fu chu. Design and Construction of bcr3/abl2 Specific Ribozyme and Further Study in Specifically Inhibiting Growth and Colony Formation of K562 Cells[J]. Chinese Journal of Biochemistry and Molecular Biology, 2000, 16(01): 10-16

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