Nuclease Stability of LNA Oligonucleotides and LNA-DNA Chimeras

Frieden, Miriam; Hansen, Henrik F.; Koch, Troels

doi:10.1081/ncn-120022731

Cited by 70 publications

(38 citation statements)

References 4 publications

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“…This bridge results in a locked 3'-endo conformation, which reduces the flexibility of the ribose, and allows for stable triplex formation [69][70][71][72]. LNA modifications within a TFO can increase the binding affinity to the target duplex DNA, and can increase resistance to digestion by nucleases [73]. Beane et al, have demonstrated that LNAs can recognize chromosomal target sequences and efficiently block endogenous expression of the progesterone and androgen receptors [74].…”

Section: Chemical Modifications Of Tfosmentioning

confidence: 99%

DNA triple helices: Biological consequences and therapeutic potential

Jain

Wang²,

Vásquez³

2008

Biochimie

262

199

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DNA structure is a critical element in determining its function. The DNA molecule is capable of adopting a variety of non-canonical structures, including three-stranded (i.e. triplex) structures, which will be the focus of this review. The ability to selectively modulate the activity of genes is a longstanding goal in molecular medicine. DNA triplex structures, either intermolecular triplexes formed by binding of an exogenously applied oligonucleotide to a target duplex sequence, or naturally occurring intramolecular triplexes (H-DNA) formed at endogenous mirror repeat sequences, present exploitable features that permit site-specific alteration of the genome. These structures can induce transcriptional repression and site-specific mutagenesis or recombination. Triplex-forming oligonucleotides (TFOs) can bind to duplex DNA in a sequence specific fashion with high affinity, and can be used to direct DNA-modifying agents to selected sequences. H-DNA plays important roles in vivo and is inherently mutagenic and recombinogenic, such that elements of the H-DNA structure may be pharmacologically exploitable. In this review we discuss the biological consequences and therapeutic potential of triple helical DNA structures. We anticipate that the information provided will stimulate further investigations aimed toward improving DNA triplexrelated gene targeting strategies for biotechnological and potential clinical applications.

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Section: Chemical Modifications Of Tfosmentioning

confidence: 99%

DNA triple helices: Biological consequences and therapeutic potential

Jain

Wang²,

Vásquez³

2008

Biochimie

262

199

View full text Add to dashboard Cite

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“…Therefore, in this study, we used third-generation LNA oligonucleotide technology, where 6 of the 16 complementary oligonucleotide residues are composed of ribose sugars that are locked in a conformation that provides much higher binding affinity to the complementary mRNA than conventional DNA and 2 0 -MOE (2 0 -O-methoxyethyl)-based oligonucleotides (10). Such LNA oligonucleotides have low single-digit nanomolar or high picomolar IC 50 values for mRNA downmodulation that have been achieved in cell culture for LNA-ASOs against AR as well as HIF-1a (13) and survivin (14,23) when used with transfection reagents.…”

Section: Discussionmentioning

confidence: 99%

“…In particular, LNA-ASOs have shown very high binding affinity to mRNA, excellent potency for target mRNA downmodulation, improved resistance to nuclease digestion, and excellent stability in plasma and tissues in preclinical studies (10). These features allow LNAASOs simplistically prepared in saline to be highly effective in vitro and in vivo (11,12).…”

Section: Introductionmentioning

confidence: 99%

Reduced Expression of the Androgen Receptor by Third Generation of Antisense Shows Antitumor Activity in Models of Prostate Cancer

Zhang

Castaneda

Dumble

et al. 2011

Molecular Cancer Therapeutics

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The androgen receptor (AR) is a member of a unique class of transcription factors because it contains a ligand-binding domain that, when activated, results in nuclear translocation and the transcriptional activation of genes associated with prostate cancer development. Although androgen deprivation therapies are effective initially for the treatment of prostate cancer, the disease eventually relapses and progresses to castrationresistant prostate cancer (CRPC). Nonetheless, the AR still plays a critical role because late-stage investigational agents that deplete testosterone (abiraterone) or block ligand binding (MDV3100) can still control tumor growth in patients with CRPC. These findings indicate that downmodulation of AR expression may provide a complementary strategy for treating CRPC. In this article, we describe a novel, locked, nucleic acid-based antisense oligonucleotide, designated EZN-4176. When administered as a single agent, EZN-4176 specifically downmodulated AR mRNA and protein, and this was coordinated with inhibition of the growth of both androgen-sensitive and CRPC tumors in vitro as well as in animal models. The effect was specific because no effect on growth was observed with a control antisense oligonucleotide that does not recognize AR mRNA, nor on tumors derived from the PC3, AR-negative, tumor cell line. In addition, EZN-4176 reduced AR luciferase reporter activity in a CRPC model derived from C4-2b cells that were implanted intratibially, indicating that the molecule may control prostate cancer that has metastasized to the bone. These data, together with the continued dependency of CRPC on the AR signaling pathway, justify the ongoing phase I evaluation of EZN-4176 in patients with CRPC.

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“…Complete stability against the 3 0 -exonuclease snake venom phosphodiesterase (SVPD) was reported for a fully modified LNA [10] while a significant increase in 3 0 -exonucleolytic stability was observed by blocking the 3 0 -end with two LNA monomers. In a study of oligonucleotide stability in serum, LNA mixmers were found to be very stable and LNA/DNA/ LNA gapmers (see section below for definition of gapmers) to be significantly more stable than DNA alone [11].…”

Section: Susceptibility Of Lna To Nucleasesmentioning

confidence: 99%