The use of 10-23 DNAzyme to selectively destroy the allele of mRNA with a unique purine-pyrimidine dinucleotide.
10-23 DNAzyme is an oligodeoxyribonucleotide-based ribonuclease. It consists of a 15-nt catalytic domain flanked by two target-specific complementary arms. It has been shown to cleave target mRNA effectively at purine (R)-pyrimidine (Y) dinucleotide. Taking advantage of this specific property, 10-23 DNAzyme was designed to cleave mRNA of a given allele at a unique RY dinucleotide while leaving the mRNA encoded from other alleles of the same gene intact. In this study, a p53-R249S (AGG-->AGT) mutant was tested. 10-23 DNAzyme was used to cut mutant mRNA at GT dinucleotide of codon 249. Both in vitro and in vivo studies showed that this DNAzyme could specifically cut the mutant p53 allele, leaving the wild-type unaffected. This proof-of-concept experiment provided a new way to knock down expression of a given allele with special single-base transversion.
Wang
Tong-Hong
TH
Institute of Biotechnology in Medicine, Department of Biotechnology and Laboratory Science in Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China.
Li
Wan-Ting
WT
Yu
Szu-Hsien
SH
Au
Lo-Chun
LC
D000483:Alleles; D045744:Cell Line, Tumor; D021881:DNA, Catalytic / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism*; D016158:Genes, p53; D006801:Humans; D009841:Oligonucleotides / Q000378:metabolism; D011687:Purines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D021881:DNA, Catalytic; D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D011687:Purines; D011743:Pyrimidines; D012333:RNA, Messenger; C402582:RNA-cleaving DNA 10-23; D016159:Tumor Suppressor Protein p53; C030986:pyrimidine; C030985:purine