Assessment of the cellular internalization of thermolytic phosphorothioate DNA oligonucleotide prodrugs

Jain, Harsh V.; Takeda, Kazuyo; Tami, Cecilia; Verthelyi, Daniela; Beaucage, Serge L.

doi:10.1016/j.bmc.2013.04.071

Cited by 11 publications

(10 citation statements)

References 37 publications

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“…The most recent data reported by Beaucage on thermosensitive PS DNA prodrugs were related to the assessment of their internalization in various cell lines [ 62 ]. The study was essentially performed with oligothymidylate models.…”

Section: Reviewmentioning

confidence: 99%

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Stimuli-responsive oligonucleotides in prodrug-based approaches for gene silencing

Debart¹,

Dupouy²,

Vasseur³

2018

Beilstein J. Org. Chem.

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Oligonucleotides (ONs) have been envisaged for therapeutic applications for more than thirty years. However, their broad use requires overcoming several hurdles such as instability in biological fluids, low cell penetration, limited tissue distribution, and off-target effects. With this aim, many chemical modifications have been introduced into ONs definitively as a means of modifying and better improving their properties as gene silencing agents and some of them have been successful. Moreover, in the search for an alternative way to make efficient ON-based drugs, the general concept of prodrugs was applied to the oligonucleotide field. A prodrug is defined as a compound that undergoes transformations in vivo to yield the parent active drug under different stimuli. The interest in stimuli-responsive ONs for gene silencing functions has been notable in recent years. The ON prodrug strategies usually help to overcome limitations of natural ONs due to their low metabolic stability and poor delivery. Nevertheless, compared to permanent ON modifications, transient modifications in prodrugs offer the opportunity to regulate ON activity as a function of stimuli acting as switches. Generally, the ON prodrug is not active until it is triggered to release an unmodified ON. However, as it will be described in some examples, the opposite effect can be sought.This review examines ON modifications in response to various stimuli. These stimuli may be internal or external to the cell, chemical (glutathione), biochemical (enzymes), or physical (heat, light). For each stimulus, the discussion has been separated into sections corresponding to the site of the modification in the nucleotide: the internucleosidic phosphate, the nucleobase, the sugar or the extremities of ONs. Moreover, the review provides a current and detailed account of stimuli-responsive ONs with the main goal of gene silencing. However, for some stimuli-responsive ONs reported in this review, no application for controlling gene expression has been shown, but a certain potential in this field could be demonstrated. Additionally, other applications in different domains have been mentioned to extend the interest in such molecules.

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Section: Reviewmentioning

confidence: 99%

“… Synthesis of thermosensitive prodrugs of ODNs containing fma thiophosphate triesters combined to positively charged 3-( N,N -dimethylamino)propyl phosphotriesters internucleoside linkages to improve cellular uptake [ 62 ]. …”

Section: Reviewmentioning

confidence: 99%

Stimuli-responsive oligonucleotides in prodrug-based approaches for gene silencing

Debart¹,

Dupouy²,

Vasseur³

2018

Beilstein J. Org. Chem.

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Section: Commentary Background Informationmentioning

confidence: 99%

“…The selection of the P(III) ester functional groups is based on cellular uptake of an uncharged thermosensitive DNA sequence, as performed previously (Jain, Takeda, Tami, Verthelyi, & Beaucage, 2013) in mammalian cells (e.g., Vero, GC-2, and HeLa). This experiment led to the discovery that replacing four neutral thiophosphate triester functionalities with four positively charged ones results in a 40-fold increase in uptake of the modified thermosensitive DNA sequence in Vero and GC-2 cells (Jain et al, 2013). Whereas the N,N-dimethylaminopropyl thiophosphate protective groups are positively charged at physiological pH, octyl thiophosphate protective groups of the thermolabile DNA sequence are hydrophobic, which is necessary to impart amphiphilicity; this property has been reported to improve in vitro cellular delivery of nucleicacid sequences (Maslov et al, 2012;Zhi et al, 2010).…”

Section: Commentary Background Informationmentioning

confidence: 99%

Synthesis, Characterization, and Function of an RNA‐Based Transfection Reagent

Jain

Boehler

Nagaraju

et al. 2018

CP Nucleic Acid Chemistry

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A synthetic 8-mer, amphipathic, trans-acting poly-2'-O-methyluridylic thiophosphate triester RNA element (2'-OMeUtaPS) can be prepared using solid-phase synthesis protocols. 2'-OMeUtaPS efficiently mediates the delivery of uncharged polyA-tailed phosphorodiamidate morpholino (PMO) sequences in HeLa pLuc 705 cells, as evidenced by flow cytometry measurements. In this cell line, 2'-OMeUtaPS-mediated transfection of an antisense polyA-tailed PMO sequence induces alternative splicing of an aberrant luciferase pre-mRNA splice site, leading to restoration of functional luciferase, as quantitatively measured using a typical luciferase assay. 2'-OMeUtaPS is also potent at delivering an uncharged antisense polyA-tailed PMO sequence in muscle cells of the mdx mouse model of muscular dystrophy; targeting the polyA-tailed PMO sequence against a splice site of the pre-mRNA encoding mutated dystrophin triggers an alternate splicing event that results in excision of the mutated exon (exon 23) from the pre-mRNA and production of functional dystrophin, as demonstrated by agarose gel electrophoresis. © 2018 by John Wiley & Sons, Inc.

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“…17 Indeed, cationic or zwitterionic ONs have shown significant cell penetrating properties and stability towards hydrolytic enzymes. [18][19][20][21][22][23][24] Particularly, the guanidinium group which has the advantage of remaining protonated over a wide pH range (pK a around 12.5) has been incorporated into ONs at selected positions on the 2 0 -OH ribose, 25,26 nucleobases 19,27 or phosphate backbone. 20 In the same way, many 2 0 -O-tethered amino groups were reported to confer nuclease resistance properties and to promote cellular uptake of such cationic ON.…”

Section: Introductionmentioning

confidence: 99%

Synthesis, binding, nuclease resistance and cellular uptake properties of 2′- O -acetalester-modified oligonucleotides containing cationic groups

Biscans¹,

Rouanet²,

Bertrand³

et al. 2015

Bioorganic & Medicinal Chemistry

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Assessment of the cellular internalization of thermolytic phosphorothioate DNA oligonucleotide prodrugs

Cited by 11 publications

References 37 publications

Stimuli-responsive oligonucleotides in prodrug-based approaches for gene silencing

Stimuli-responsive oligonucleotides in prodrug-based approaches for gene silencing

Synthesis, Characterization, and Function of an RNA‐Based Transfection Reagent

Synthesis, binding, nuclease resistance and cellular uptake properties of 2′- O -acetalester-modified oligonucleotides containing cationic groups

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