Limits...
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

Delivery of a cell-membrane-impermeable dirutheniumcomplex tolive cancer cells by using the GNP–M1/MC2(EG12)3. (A) Chemical structure of the diruthenium(II) complex, BPY.(B) Confocal phase contrast (left), fluorescence (middle), and mergedoptical/fluorescence images (right) of HeLa cells after treatmentwith the BPY for 3 h at 37 °C. (C) Confocal phase contrast (left),fluorescence (middle), and merged optical/fluorescence (right) imagesof HeLa cells after incubation with GNP–M1/MC2(EG12)3–BPY for 3 h at 37 °C.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4554298&req=5

fig6: Delivery of a cell-membrane-impermeable dirutheniumcomplex tolive cancer cells by using the GNP–M1/MC2(EG12)3. (A) Chemical structure of the diruthenium(II) complex, BPY.(B) Confocal phase contrast (left), fluorescence (middle), and mergedoptical/fluorescence images (right) of HeLa cells after treatmentwith the BPY for 3 h at 37 °C. (C) Confocal phase contrast (left),fluorescence (middle), and merged optical/fluorescence (right) imagesof HeLa cells after incubation with GNP–M1/MC2(EG12)3–BPY for 3 h at 37 °C.

Mentions: To demonstrate the general use of the GNP–M1/MC2(EG12)3 for intracellular delivery of other types ofDNA binding agents, we have further employed it to deliver a fluorescentdiruthenium(II) complex, [(bpy)2Ru(tpphz)Ru(bpy)2]4+, denoted as BPY (Figure 6A). Unlike DOX and PI molecules which bindto DNA mainly through intercalation, BPY is a DNA groove binder.60 BPY has been shown to be impermeable to livecell membranes and therefore cannot enter cells on its own.60 This property is further confirmed from ourresults shown in Figure 6B: 3 h incubation of free BPY with HeLa cells produces negligibleBPY fluorescence inside the cells, suggesting no significant celluptake. In contrast, incubation of HeLa cells with the BPY mixed withthe GNP–M1/MC2(EG12)3 for 3 h yieldsstrong BPY fluorescence inside HeLa cells, suggesting that the GNP–M1/MC2(EG12)3 can effectively carry the BPY molecules andsuccessfully deliver them into live HeLa cells. Together, these resultsdemonstrate that the GNP–M1/MC2(EG12)3 reported herein has great potential for intracellular delivery ofa wide range of DNA-intercalating agents. Its excellent stabilityand resistance against nonspecific adsorption and enzymatic degradation,together with high cell uptake, should make it an effective nanocarrierfor intracellular delivery of any DNA-binding/intercalating reagents.Given a large number of drug molecules and metal complexes are knownto be DNA-binders,61 the robust, versatilePEGylated DNA–GNP nanocarrier reported herein should have broadapplications in bioimaging, drug delivery, and therapeutics, possiblyeven at the in vivo level.


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)

Delivery of a cell-membrane-impermeable dirutheniumcomplex tolive cancer cells by using the GNP–M1/MC2(EG12)3. (A) Chemical structure of the diruthenium(II) complex, BPY.(B) Confocal phase contrast (left), fluorescence (middle), and mergedoptical/fluorescence images (right) of HeLa cells after treatmentwith the BPY for 3 h at 37 °C. (C) Confocal phase contrast (left),fluorescence (middle), and merged optical/fluorescence (right) imagesof HeLa cells after incubation with GNP–M1/MC2(EG12)3–BPY for 3 h at 37 °C.
© Copyright Policy
Related In: Results  -  Collection

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

fig6: Delivery of a cell-membrane-impermeable dirutheniumcomplex tolive cancer cells by using the GNP–M1/MC2(EG12)3. (A) Chemical structure of the diruthenium(II) complex, BPY.(B) Confocal phase contrast (left), fluorescence (middle), and mergedoptical/fluorescence images (right) of HeLa cells after treatmentwith the BPY for 3 h at 37 °C. (C) Confocal phase contrast (left),fluorescence (middle), and merged optical/fluorescence (right) imagesof HeLa cells after incubation with GNP–M1/MC2(EG12)3–BPY for 3 h at 37 °C.
Mentions: To demonstrate the general use of the GNP–M1/MC2(EG12)3 for intracellular delivery of other types ofDNA binding agents, we have further employed it to deliver a fluorescentdiruthenium(II) complex, [(bpy)2Ru(tpphz)Ru(bpy)2]4+, denoted as BPY (Figure 6A). Unlike DOX and PI molecules which bindto DNA mainly through intercalation, BPY is a DNA groove binder.60 BPY has been shown to be impermeable to livecell membranes and therefore cannot enter cells on its own.60 This property is further confirmed from ourresults shown in Figure 6B: 3 h incubation of free BPY with HeLa cells produces negligibleBPY fluorescence inside the cells, suggesting no significant celluptake. In contrast, incubation of HeLa cells with the BPY mixed withthe GNP–M1/MC2(EG12)3 for 3 h yieldsstrong BPY fluorescence inside HeLa cells, suggesting that the GNP–M1/MC2(EG12)3 can effectively carry the BPY molecules andsuccessfully deliver them into live HeLa cells. Together, these resultsdemonstrate that the GNP–M1/MC2(EG12)3 reported herein has great potential for intracellular delivery ofa wide range of DNA-intercalating agents. Its excellent stabilityand resistance against nonspecific adsorption and enzymatic degradation,together with high cell uptake, should make it an effective nanocarrierfor intracellular delivery of any DNA-binding/intercalating reagents.Given a large number of drug molecules and metal complexes are knownto be DNA-binders,61 the robust, versatilePEGylated DNA–GNP nanocarrier reported herein should have broadapplications in bioimaging, drug delivery, and therapeutics, possiblyeven at the in vivo level.

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