The stability of different forms of double-stranded decoy DNA in serum and nuclear extracts

Chu, Barbara C.F.; Orgel, Leslie E.

doi:10.1093/nar/20.21.5857

Cited by 77 publications

(82 citation statements)

References 9 publications

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“…[9][10][11][12] Dumbbell-type ODN is reported to increase nuclease resistance and uptake into cells, compared to the chemically modified linear ODN. 28,29 Accordingly, we designed a novel circular dumbbell ODN containing two binding sites for E2F in a single decoy molecule without an open end, thus allowing the multiple targeting of a target promoter site or more than one promoter site. As expected, CD-E2F ODN was more stable than PS-E2F ODN in vitro as well as in vivo.…”

Section: Discussionmentioning

confidence: 99%

Novel E2F decoy oligodeoxynucleotides inhibit in vitro vascular smooth muscle cell proliferation and in vivo neointimal hyperplasia

Morishita

Kaneda

et al. 2002

Gene Ther

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The transcription factor, E2F, plays a critical role in the transactivation of several genes involved in cell cycle regulation. Previous studies showed that the transfection of cis element double-stranded decoy oligodeoxynucleotides (ODNs) corresponding to E2F binding sites inhibited the proliferation of vascular smooth muscle cells (VSMCs) and neointimal hyperplasia in injured vessels. We have developed a novel E2F decoy ODN with a circular dumbbell structure (CD-E2F) and compared its effects with those of the conventional phosphorothioated E2F decoy (PS-E2F) ODN. CD-E2F ODN was more stable than PS-E2F ODN, largely preserving its structural integrity after incubation in the presence of nucleases and sera. Moreover, CD-E2F ODN inhibited high glucose-and serum-induced transcriptional expression of cell cycle regulatory genes more strongly than PS-E2F ODN. Transfection of CD-E2F ODN resulted in more effective inhibition of VSMC proliferation in vitro and neointimal formation in vivo, compared with PS-E2F ODN. An approximately 40-50% lower dose of CD-E2F ODN than PS-E2F ODN was sufficient to attain similar effects. In conclusion, our results indicate that CD-E2F ODN may be a valuable tool in gene therapy protocols for inhibiting VSMC proliferation and studying transcriptional regulation.

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Section: Discussionmentioning

confidence: 99%

Novel E2F decoy oligodeoxynucleotides inhibit in vitro vascular smooth muscle cell proliferation and in vivo neointimal hyperplasia

Morishita

Kaneda

et al. 2002

Gene Ther

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“…When DNA with only a phosphodiester backbone was formed into a dumbbell-like structure, it certainly displayed resistance to DNase. [49][50][51] Schmidt et al 52 synthesized an ODN with a phosphodiester backbone in a dumbbell-like structure designed to include CpG motifs in hairpin-loops ( Figure 3A). By utilizing the actions of peripheral blood mononuclear cells, immunostimulatory effects similar to linear-structure CpG ODNs with phosphorothioate backbones at both ends were obtained.…”

Section: Synthetic Cpg Odns With Various Structures and Their Immunosmentioning

confidence: 99%

Structure-dependent immunostimulatory effect of CpG oligodeoxynucleotides and their delivery system

Hanagata¹

2012

IJN

162

139

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Unmethylated cytosine-phosphate-guanosine (CpG) oligodeoxynucleotides (ODNs) are recognized by Toll-like receptor 9 (TLR9) found in antigen-presenting cells and B cells and can activate the immune system. Using CpG ODNs as an adjuvant has been found to be effective for treating infectious diseases, cancers, and allergies. Because natural ODNs with only a phosphodiester backbone are easily degraded by nuclease (deoxyribonuclease [DNase]) in serum, CpG ODNs with a phosphorothioate backbone have been studied for clinical application. CpG ODNs with a phosphorothioate backbone have raised concern regarding undesirable side effects; however, several CpG ODNs with only a phosphodiester backbone have been reported to be stable in serum and to show an immunostimulatory effect. In recent years, research has been conducted on delivery systems for CpG ODNs using nanoparticles (NPs). The advantages of NP-based delivery of CpG ODN include (1) it can protect CpG ODN from DNase, (2) it can retain CpG ODN inside the body for a long period of time, (3) it can improve the cellular uptake efficiency of CpG ODN, and (4) it can deliver CpG ODN to the target tissues. Because the target cells of CpG ODN are cells of the immune system and TLR9, the receptor of CpG ODN is localized in endolysosomes, CpG ODN delivery systems are required to have qualities different from other nucleic acid drugs such as antisense DNA and small interfering RNA. Studies until now have reported various NPs as carriers for CpG ODN delivery. This review presents DNase-resistant CpG ODNs with various structures and their immunostimulatory effects and also focuses on delivery systems of CpG ODNs that utilize NPs. Because CpG ODNs interact with TLR9 and activate both the innate and the adaptive immune system, the application of CpG ODNs for the treatment of cancers, infectious diseases, and allergies holds great promise.

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“…In order to protect the free end of the linear DNA from exonuclease degradation, it may be helpful to form a covalently closed end using oligonucleotides ("dumbbell DNA") [33], or to otherwise modify the free end by attachment to a peptide or modification of the nucleotides at this end. Additional DNA elements, such as polyadenylation signals, will also likely be helpful in eukaryotic cells.…”

Section: Evaluation Of the Hypothesismentioning

confidence: 99%

Method of Detecting and Targeting Mutations in Cancer

Edwards

2007

Nature Precedings

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While there are many differences between tumor and non-tumor cells, the basic underlying distinction is in the DNA. Tumor cells harbor mutations, at least some of which are not present in non-tumor cells. Thus, a method of directly targeting cells containing specific mutations has potential for detection or treatment of cancer without the toxicity associated with more indirect approaches. Also, as mutations are a necessary component of malignancy, such a method is potentially applicable to all tumors. And, as a consequence of the molecules binding to the mutation, the cells will be destroyed. (3) Deliver these molecules to all cells (or at least all tumor cells). I hypothesize that such molecules can now be constructed using sequence-specific DNA binding proteins (such as customized zinc-finger DNA binding proteins) fused to transcriptional activator domains (such as VP16) and reporter or toxin genes. The necessary genes can be linked to the DNA binding proteins utilizing a recently described method based on expressed protein ligation.

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The stability of different forms of double-stranded decoy DNA in serum and nuclear extracts

Cited by 77 publications

References 9 publications

Novel E2F decoy oligodeoxynucleotides inhibit in vitro vascular smooth muscle cell proliferation and in vivo neointimal hyperplasia

Novel E2F decoy oligodeoxynucleotides inhibit in vitro vascular smooth muscle cell proliferation and in vivo neointimal hyperplasia

Structure-dependent immunostimulatory effect of CpG oligodeoxynucleotides and their delivery system

Method of Detecting and Targeting Mutations in Cancer

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