Design and kinetic analysis of hammerhead ribozyme and DNAzyme that specifically cleave TEL-AML1 chimeric mRNA

Biochem Biophys Res Commun. 2008 Sep 12;374(1):169-74. doi: 10.1016/j.bbrc.2008.07.008. Epub 2008 Jul 15.

Abstract

In order to develop the oligonucleotides to abolish an expression of TEL-AML1 chimeric RNA, which is a genetic aberration that causes the acute lymphoblastic leukemia (ALL), hammerhead ribozymes and deoxyoligoribozymes that can specifically cleave TEL-AML1 fusion RNA were designed. Constructs of the deoxyribozyme with an asymmetric substrate binding arm (Dz26) and the hammerhead ribozyme with a 4nt-bulged substrate binding arm in the stem III (buRz28) were able to cleave TEL-AML1 chimeric RNA specifically at sites close to the junction in vitro, without cleaving the normal TEL and AML1 RNA. Single-turnover kinetic analysis under enzyme-excess condition revealed that the buRz28 is superior to the Dz26 in terms of substrate binding and RNA-cleavage. In conjunction with current progress in a gene-delivery technology, the designed oligonucleotides that specifically cleave the TEL-AML1 chimeric mRNA are hoped to be applicable for the treatment of ALL in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Cell Line, Tumor
  • Core Binding Factor Alpha 2 Subunit / antagonists & inhibitors*
  • Core Binding Factor Alpha 2 Subunit / genetics
  • DNA, Catalytic / chemistry
  • DNA, Catalytic / genetics
  • DNA, Catalytic / metabolism*
  • Gene Transfer Techniques
  • Humans
  • Kinetics
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism*
  • Oncogene Proteins, Fusion / antagonists & inhibitors*
  • Oncogene Proteins, Fusion / genetics
  • RNA, Catalytic / chemistry
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism*

Substances

  • Core Binding Factor Alpha 2 Subunit
  • DNA, Catalytic
  • Oligonucleotides
  • Oncogene Proteins, Fusion
  • RNA, Catalytic
  • RNA, Messenger
  • TEL-AML1 fusion protein
  • hammerhead ribozyme