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Terminal PEGylated DNA-Gold Nanoparticle Conjugates Offering High Resistance to Nuclease Degradation and Efficient Intracellular Delivery of DNA Binding Agents.

Song L, Guo Y, Roebuck D, Chen C, Yang M, Yang Z, Sreedharan S, Glover C, Thomas JA, Liu D, Guo S, Chen R, Zhou D - ACS Appl Mater Interfaces (2015)

Bottom Line: A barrier limiting its in vivo effectiveness is limited resistance to nuclease degradation and nonspecific interaction with blood serum contents.The PEGylated DNA-GNP conjugate still retains a high cell uptake property, making it an attractive intracellular delivery nanocarrier for DNA binding reagents.Moreover, it can be used for efficient delivery of some cell-membrane-impermeable reagents such as propidium iodide (a DNA intercalating fluorescent dye currently limited to the use of staining dead cells only) and a diruthenium complex (a DNA groove binder), for successful staining of live cells.

View Article: PubMed Central - PubMed

Affiliation: School of Chemistry and Astbury Structure for Molecular Biology, University of Leeds , Leeds LS2 9JT, U.K.

ABSTRACT
Over the past 10 years, polyvalent DNA-gold nanoparticle (DNA-GNP) conjugate has been demonstrated as an efficient, universal nanocarrier for drug and gene delivery with high uptake by over 50 different types of primary and cancer cell lines. A barrier limiting its in vivo effectiveness is limited resistance to nuclease degradation and nonspecific interaction with blood serum contents. Herein we show that terminal PEGylation of the complementary DNA strand hybridized to a polyvalent DNA-GNP conjugate can eliminate nonspecific adsorption of serum proteins and greatly increases its resistance against DNase I-based degradation. The PEGylated DNA-GNP conjugate still retains a high cell uptake property, making it an attractive intracellular delivery nanocarrier for DNA binding reagents. We show that it can be used for successful intracellular delivery of doxorubicin, a widely used clinical cancer chemotherapeutic drug. Moreover, it can be used for efficient delivery of some cell-membrane-impermeable reagents such as propidium iodide (a DNA intercalating fluorescent dye currently limited to the use of staining dead cells only) and a diruthenium complex (a DNA groove binder), for successful staining of live cells.

No MeSH data available.


Related in: MedlinePlus

(A) Schematic procedures of our approach toPEGylated DNA–GNPdrug nanocarriers. Thiolated PR-DNA (denoted as M1) was first loadedonto a citrate-stabilized 14 nm GNP via gold–thiol self-assemblyto form GNP–M1, which was then hybridized to complementaryMC2 (unmodified, route 1) or PEG-modified MC2s (route 2) to form theGNP–M1/MC2(PEG) carriers. (B) Schematic of MC2(EG12)3 preparation via the Michael addition between the maleimide-modifiedthree-chain oligo(ethylene glycol) and the MC2-free sulfhydryl group,forming a stable covalently linked MC2(EG12)3.
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fig1: (A) Schematic procedures of our approach toPEGylated DNA–GNPdrug nanocarriers. Thiolated PR-DNA (denoted as M1) was first loadedonto a citrate-stabilized 14 nm GNP via gold–thiol self-assemblyto form GNP–M1, which was then hybridized to complementaryMC2 (unmodified, route 1) or PEG-modified MC2s (route 2) to form theGNP–M1/MC2(PEG) carriers. (B) Schematic of MC2(EG12)3 preparation via the Michael addition between the maleimide-modifiedthree-chain oligo(ethylene glycol) and the MC2-free sulfhydryl group,forming a stable covalently linked MC2(EG12)3.

Mentions: To address the problem of serum protein nonspecific adsorption,the Mirkin group used a post-treatment of the formed DNA–GNPwith a thiolated poly(ethylene glycol) (PEG). Despite success, a drawbackwas a reduced DNA/RNA loading on the GNP, due to competitive displacementof the thiolated nucleic acid strands on the GNP surface by the thiolatedPEG passivation molecules. As a result, the number of functional DNA/RNAstrands on each ∼14 nm GNP was found to be only ∼35,18 a considerable reduction from the typical ≥100strands found for nontreated DNA–GNPs.1−12 Herein we report a new PEGylation strategy for the DNA–GNPvia terminal PEGylation of the complementary strand (MC2). The specifichybridization between the PR-DNA–GNP and MC2(PEG) then completesthe carrier PEGylation (Figure 1A). An advantage of this strategy over the post-thiolatedPEG treatment is that it yields more functional DNA strands per GNP(ca. 110 vs 35), making it potentially a more effective drug or genenanocarrier. We show that our PEGylation approach offers completeresistance to nonspecific adsorption of serum proteins in cell culturemedia and provides >10 times higher resistance to DNase I-mediatedenzymatic digestion. Moreover, the PEGylated DNA–GNP nanocarrierstill retains high cell uptake which can be exploited for efficientdelivery of both chemotherapeutic drugs (ca. DOX) and some cell membrane-impermeablereagents to live cells.


Terminal PEGylated DNA-Gold Nanoparticle Conjugates Offering High Resistance to Nuclease Degradation and Efficient Intracellular Delivery of DNA Binding Agents.

Song L, Guo Y, Roebuck D, Chen C, Yang M, Yang Z, Sreedharan S, Glover C, Thomas JA, Liu D, Guo S, Chen R, Zhou D - ACS Appl Mater Interfaces (2015)

(A) Schematic procedures of our approach toPEGylated DNA–GNPdrug nanocarriers. Thiolated PR-DNA (denoted as M1) was first loadedonto a citrate-stabilized 14 nm GNP via gold–thiol self-assemblyto form GNP–M1, which was then hybridized to complementaryMC2 (unmodified, route 1) or PEG-modified MC2s (route 2) to form theGNP–M1/MC2(PEG) carriers. (B) Schematic of MC2(EG12)3 preparation via the Michael addition between the maleimide-modifiedthree-chain oligo(ethylene glycol) and the MC2-free sulfhydryl group,forming a stable covalently linked MC2(EG12)3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4554298&req=5

fig1: (A) Schematic procedures of our approach toPEGylated DNA–GNPdrug nanocarriers. Thiolated PR-DNA (denoted as M1) was first loadedonto a citrate-stabilized 14 nm GNP via gold–thiol self-assemblyto form GNP–M1, which was then hybridized to complementaryMC2 (unmodified, route 1) or PEG-modified MC2s (route 2) to form theGNP–M1/MC2(PEG) carriers. (B) Schematic of MC2(EG12)3 preparation via the Michael addition between the maleimide-modifiedthree-chain oligo(ethylene glycol) and the MC2-free sulfhydryl group,forming a stable covalently linked MC2(EG12)3.
Mentions: To address the problem of serum protein nonspecific adsorption,the Mirkin group used a post-treatment of the formed DNA–GNPwith a thiolated poly(ethylene glycol) (PEG). Despite success, a drawbackwas a reduced DNA/RNA loading on the GNP, due to competitive displacementof the thiolated nucleic acid strands on the GNP surface by the thiolatedPEG passivation molecules. As a result, the number of functional DNA/RNAstrands on each ∼14 nm GNP was found to be only ∼35,18 a considerable reduction from the typical ≥100strands found for nontreated DNA–GNPs.1−12 Herein we report a new PEGylation strategy for the DNA–GNPvia terminal PEGylation of the complementary strand (MC2). The specifichybridization between the PR-DNA–GNP and MC2(PEG) then completesthe carrier PEGylation (Figure 1A). An advantage of this strategy over the post-thiolatedPEG treatment is that it yields more functional DNA strands per GNP(ca. 110 vs 35), making it potentially a more effective drug or genenanocarrier. We show that our PEGylation approach offers completeresistance to nonspecific adsorption of serum proteins in cell culturemedia and provides >10 times higher resistance to DNase I-mediatedenzymatic digestion. Moreover, the PEGylated DNA–GNP nanocarrierstill retains high cell uptake which can be exploited for efficientdelivery of both chemotherapeutic drugs (ca. DOX) and some cell membrane-impermeablereagents to live cells.

Bottom Line: A barrier limiting its in vivo effectiveness is limited resistance to nuclease degradation and nonspecific interaction with blood serum contents.The PEGylated DNA-GNP conjugate still retains a high cell uptake property, making it an attractive intracellular delivery nanocarrier for DNA binding reagents.Moreover, it can be used for efficient delivery of some cell-membrane-impermeable reagents such as propidium iodide (a DNA intercalating fluorescent dye currently limited to the use of staining dead cells only) and a diruthenium complex (a DNA groove binder), for successful staining of live cells.

View Article: PubMed Central - PubMed

Affiliation: School of Chemistry and Astbury Structure for Molecular Biology, University of Leeds , Leeds LS2 9JT, U.K.

ABSTRACT
Over the past 10 years, polyvalent DNA-gold nanoparticle (DNA-GNP) conjugate has been demonstrated as an efficient, universal nanocarrier for drug and gene delivery with high uptake by over 50 different types of primary and cancer cell lines. A barrier limiting its in vivo effectiveness is limited resistance to nuclease degradation and nonspecific interaction with blood serum contents. Herein we show that terminal PEGylation of the complementary DNA strand hybridized to a polyvalent DNA-GNP conjugate can eliminate nonspecific adsorption of serum proteins and greatly increases its resistance against DNase I-based degradation. The PEGylated DNA-GNP conjugate still retains a high cell uptake property, making it an attractive intracellular delivery nanocarrier for DNA binding reagents. We show that it can be used for successful intracellular delivery of doxorubicin, a widely used clinical cancer chemotherapeutic drug. Moreover, it can be used for efficient delivery of some cell-membrane-impermeable reagents such as propidium iodide (a DNA intercalating fluorescent dye currently limited to the use of staining dead cells only) and a diruthenium complex (a DNA groove binder), for successful staining of live cells.

No MeSH data available.


Related in: MedlinePlus