Inhibition of Human Papillomavirus Expression Using DNAzymes

Abstract

Deoxyribozymes (DXZs) are catalytic oligodeoxynucleotides capable of performing diverse functions including the specific cleavage of a target RNA. These molecules represent a new type of therapeutic oligonucleotides combining the efficiency of ribozymes and the intracellular endurance and simplicity of modified antisense oligonucleotides. Commonly used DXZs include the 8-17 and 10-23 motifs, which have been engineered to destroy disease-associated genes with remarkable efficiency. Targeting DXZs to disease-associated transcripts requires extensive biochemical testing to establish target RNA accessibility, catalytic efficiency, and nuclease sensibility. The usage of modified nucleotides to render nuclease-resistance DXZs must be counterweighted against deleterious consequences on catalytic activity. Further intracellular testing is required to establish the effect of microenvironmental conditions on DXZ activity and off-target issues. Application of modified DXZs to cervical cancer results in specific growth inhibition, cell death, and apoptosis. Thus, DXZs represent a highly effective antisense moiety with minimal secondary effects.

Therapeutic startegies for human papillomavirus infection and associated cancers

“…Because of deleterious off-target effects in intracellular milieu of an otherwise specific and proven catalytic molecule against HPV in biochemical assays, the pursuit to make this class of molecules as potential therapeutics faded considerably. To overcome problems encountered with ribozymes, alternative molecules particularly the DNAzymes were explored (123,124). DNAzymes combined the efficiency of ribozymes and intracellular stability and simplicity of AS-ODNs (125).…”

“…DNAzymes combined the efficiency of ribozymes and intracellular stability and simplicity of AS-ODNs (125). Out of the two commonly used DNAzymes, a widely used and well-characterized 10-23 motif has been engineered to destroy HPV oncogenes (123,124). DNAzyme Dz1023-434 showed efficient cleavage against HPV-16 E6/E7 mRNA within bona fide antisense window at nt 410-445 even in low (Mg ++ ) condition cell free systems.…”

Fluorescence Visualization Screening for EBV‐LMP1‐Targeted DNAzymes

“…8 Nowadays, the 10-23 DNAzymes have been widely employed to inhibit protein expression in experiments. 9,10 Recent studies have demonstrated that LMP1 expression can be suppressed by LMP1-targeted DNAzymes, which stimulate cell apoptosis, 11,12 inhibit cell proliferation and xenograft growth in nude mice, 13 and enhance radiosensitivity in NPC. 14 DNAzymes feature design simplicity, target-specific cleavage, serum stability, and catalytic activity.…”

“…Deoxyribozymes (DNAzymes, DZs) are synthetic single‐stranded DNA catalysts that can be bound to their complementary sequences in a target messenger RNA (mRNA) by Watson‐Crick base pairing and cleave the mRNA with predetermined phosphodiester linkages 8 . Nowadays, the 10‐23 DNAzymes have been widely employed to inhibit protein expression in experiments 9,10 . Recent studies have demonstrated that LMP1 expression can be suppressed by LMP1‐targeted DNAzymes, which stimulate cell apoptosis, 11,12 inhibit cell proliferation and xenograft growth in nude mice, 13 and enhance radiosensitivity in NPC 14 .…”