Double-tailed lipid modification as a promising candidate for oligonucleotide delivery in mammalian cells

Ugarte‐Uribe, Begoña; Grijalvo, Santiago; Busto, Jon V.; Martı́n, César; Eritja, Ramón; Goñi, Félix M.; Alkorta, Itziar

doi:10.1016/j.bbagen.2013.06.013

Cited by 12 publications

(19 citation statements)

References 65 publications

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“…A recent study showed that double tailed, saturated lipid conjugated to an oligonucleotide significantly improved cellular uptake in a variety of tumor cell types (Ugarte-Uribe et al, 2013). Impressively, the conjugate significantly surpassed the cellular uptake achieved by transfection in HeLa cells.…”

Section: Lipid Conjugatesmentioning

confidence: 99%

Cellular Uptake and Intracellular Trafficking of Oligonucleotides: Implications for Oligonucleotide Pharmacology

Juliano

Xin

Carver

et al. 2014

Nucleic Acid Therapeutics

110

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One of the major constraints on the therapeutic use of oligonucleotides is inefficient delivery to their sites of action in the cytosol or nucleus. Recently it has become evident that the pathways of cellular uptake and intracellular trafficking of oligonucleotides can strongly influence their pharmacological actions. Here we provide background information on the basic processes of endocytosis and trafficking and then review recent literature on targeted delivery and subcellular trafficking of oligonucleotides in that context. A variety of approaches including molecular scale ligand-oligonucleotide conjugates, ligand-targeted nanocarriers, and the use of small molecules to enhance oligonucleotide effects are discussed.

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Section: Lipid Conjugatesmentioning

confidence: 99%

Cellular Uptake and Intracellular Trafficking of Oligonucleotides: Implications for Oligonucleotide Pharmacology

Juliano

Xin

Carver

et al. 2014

Nucleic Acid Therapeutics

110

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“…SPB preparation was performed as described previously. 23 Sucrose-enclosing GUVs were prepared using the electroformation method as described previously. 23 DOPC, DOPC/eSM/Chol (2:2:1) and eSM:Chol (2:1) stock solutions were prepared containing 0.4 mol% DiD (Invitrogen, Waltham, Massachusetts, USA).…”

Section: Spb and Guv Preparationmentioning

confidence: 99%

“…23 Sucrose-enclosing GUVs were prepared using the electroformation method as described previously. 23 DOPC, DOPC/eSM/Chol (2:2:1) and eSM:Chol (2:1) stock solutions were prepared containing 0.4 mol% DiD (Invitrogen, Waltham, Massachusetts, USA). Vesicles were generated in a 300 mM sucrose solution at 60ºC, a temperature rendering bilayers in a fluid phase state.…”

Section: Spb and Guv Preparationmentioning

confidence: 99%

“…30 Furthermore, to carry out membrane binding studies and cellular uptake experiments, fluorescent dye Alexa Fluor 488 was introduced into the LOC-20 model conjugate at 3'-termini (LOC-21) taking advantage of the presence of the free primary amine group, as previously described. 23 Although model membranes cannot mimic the complex cellular environment, their use would allow the study of certain biophysical aspects of oligonucleotide delivery under controlled experimental conditions and welldefined lipid compositions. 34,35 We could clearly observe that this LOC could be efficiently inserted into DOPC/eSM/Chol (2:2:1) monolayers at initial π above 35 mN/m ( Figure S5A in Supporting Information), indicating that its incorporation into different domains of cell membranes would be expected since biological membranes are considered to support a lateral pressure π ≈30 mN/m, as previously observed in several cell lines, 23 albeit with large fluctuations around this average value.…”

Section: Insertion Into Model Membranesmentioning

confidence: 99%

“…23 Although model membranes cannot mimic the complex cellular environment, their use would allow the study of certain biophysical aspects of oligonucleotide delivery under controlled experimental conditions and welldefined lipid compositions. 34,35 We could clearly observe that this LOC could be efficiently inserted into DOPC/eSM/Chol (2:2:1) monolayers at initial π above 35 mN/m ( Figure S5A in Supporting Information), indicating that its incorporation into different domains of cell membranes would be expected since biological membranes are considered to support a lateral pressure π ≈30 mN/m, as previously observed in several cell lines, 23 albeit with large fluctuations around this average value. 36 These results were in agreement with what was previously observed in GUVs and SPBs composed of the same ternary mixture, 23 where the insertion of LOC showed a homogeneous incorporation into both liquid-disordered (L d ) and liquid-ordered (L o ) domains.…”

Section: Insertion Into Model Membranesmentioning

confidence: 99%

See 2 more Smart Citations

Lipid-modified oligonucleotide conjugates: Insights into gene silencing, interaction with model membranes and cellular uptake mechanisms

Ugarte‐Uribe

Grijalvo

Núñez-Pertíñez

et al. 2017

Bioorganic & Medicinal Chemistry

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The ability of oligonucleotides to silence specific genes or inhibit the biological activity of specific proteins has generated great interest in their use as research tools and therapeutic agents. Unfortunately, their biological applications meet the limitation of their poor cellular accessibility. Developing an appropriate delivery system for oligonucleotides is essential to achieve their efficient cellular uptake. In the present work a series of phosphorothioate lipid-oligonucleotide hybrids were synthesized introducing covalently single or double lipid tails at both 3'- and 5'-termini of an antisense oligonucleotide. Gene transfections in cultured cells showed antisense luciferase inhibition without the use of a transfecting agent for conjugates modified with the double-lipid tail at 5'-termini. The effect of the double lipid-tailed modification was further studied in detail in several model membrane systems as well as in cellular uptake experiments. During these studies the spontaneous formation of self-assembled microstructures is clearly observed. Lipidation allowed the efficient incorporation of the oligonucleotide in HeLa cells by a macropinocytosis mechanism without causing cytotoxicity in cells or altering the binding properties of the oligonucleotide conjugates. In addition, both single- and double-tailed compounds showed a similar behavior in lipid model membranes, making them useful in nucleotide-based technologies.

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Chemical Modifications in Nucleic Acids for Therapeutic and Diagnostic Applications

Fàbrega

Aviñó

Eritja

2021

The Chemical Record

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The last decade has witnessed the blooming of nucleic acids for therapeutic and diagnostic applications. In the present article, we describe the most important results from our group in this area covering the international context that surrounded this research. These include the study of modifications at the terminal and internal positions of siRNA duplexes to enhance nuclease resistance, increase loading of the antisense strand to RISC and avoid side effects such as activation of immune response and sense strand misloading. Then, we describe the design of novel lipid, carbohydrate and peptide conjugates to enhance cellular uptake. Finally, we describe the use of nanostructures for drug delivery and for the controlled deposition of matter on surfaces. We invite the readers to submerge into a highly interdisciplinary discipline that combines organic chemistry, biochemical assays, pharmacology issues as well as materials chemistry and structural studies in order to increase the applications of nucleic acids.

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Double-tailed lipid modification as a promising candidate for oligonucleotide delivery in mammalian cells

Abstract: Compared to the commercial lipoplex approach, oligodeoxyribonucleotide conjugation with C28 dual lipid modification seems to be promising to improve oligonucleotide delivery in mammalian cells.

Cited by 12 publications

References 65 publications

Cellular Uptake and Intracellular Trafficking of Oligonucleotides: Implications for Oligonucleotide Pharmacology

Cellular Uptake and Intracellular Trafficking of Oligonucleotides: Implications for Oligonucleotide Pharmacology

Lipid-modified oligonucleotide conjugates: Insights into gene silencing, interaction with model membranes and cellular uptake mechanisms

Chemical Modifications in Nucleic Acids for Therapeutic and Diagnostic Applications

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