Real-Time Fluorescence Tracking of Gene Delivery via Multifunctional Nanocomposites

Abstract

Fluorescence imaging of transduced cells and tissues is valuable in the development of gene vectors and the evaluation of gene therapy efficacy. We report here the simple and rational design of multifunctional nanocomposites (NCs) for simultaneous gene delivery and fluorescence tracking based on ZnS:Mn(2+) quantum dots (QDs) and positively charged polymer coating. The positively charged imidazole in the as-synthesized amphiphilic copolymer can be used for gene loading via electrostatic interaction. While the introduced poly(ethylene glycol) (PEG) can be used to reduce the binding of plasma proteins to nanovectors and minimize clearance by the reticuloendothelial system after intravenous administration. Most importantly, these multifunctional nanovectors showed much lower cellular toxicity than the commercial polyethylenimine (PEI) transfection vectors. On the basis of the red fluorescence of QDs, we can real-time track the gene delivery in cells, and the transfection efficacy of pDNA encoding enhanced green fluorescence protein (pEGFP) was monitored via the green fluorescence of the GFP expressed by the pDNA delivered into the nuclei. Fluorescence imaging analysis confirmed that the QDs-based nanovectors delivered pDNA into HepG2 cells efficiently. These new insights and capabilities pave a new way toward nanocomposite engineering for fluorescence imaging tracking of gene therapy.

Upconversion luminescence tracking of gene delivery via multifunctional nanocapsules

“…Then newly synthesized multifunctional amphiphilic polymer(MFAP), together with polyethylene glycol-poly (lactic-co-glycolic acid) (PEG-PLGA), was further employed to coat hydrophobic NaYF 4 :Yb 3+ /Er 3+ UCNPs, affording UCNPs@MFAP nanocapsules with high water dispersibility, biocompatibility and low cytotoxicity due to the introduction of PEG moiety. According to previous work, (Bai et al 2014) PEG-PLGA could reduce nonspecific absorption of plasma proteins, prolong the in vivo circulation time and facilitate accumulation targeted tissues. (Pozzi et al 2014;Wonganan et al 2011) Besides, the hydrophobic moiety PLGA could facilitate the encapsulation of hydrophobic UCNPs, which Scheme1.…”

Luminescent nanocarriers for simultaneous drug or gene delivery and imaging tracking

“…Our group [22] recently constructed inorganic (ZnS:Mn 2+ QDs)-organic (positively-charged amphiphilic polymer) multifunctional nanovectors to illuminate the pathway of plasmid DNA (pDNA for green fluorescence proteins) into HepG2 cells ( Fig. 3a and 3b).…”

“…The combination of different therapies, such as photothermal therapy, photodynamic therapy and chemotherapy in one nanocapsule is challenging. {Reprinted with permission from [22]; ©2014 American Chemical Society}. …”

“…QDs, with unique tunable optical properties, have frequently been utilized for designing multifunctional drug-or gene-delivery nanovectors [22][23][24]. Lanthanide-doped nanocrystals, due to their upconversion luminescence (UCL) properties [16,20,[25][26][27], have become popular candidates especially in in vivo tracking studies.…”

Intracellular Delivery by Shape Anisotropic Magnetic Particle–Induced Cell Membrane Cuts

“…[5][6][7] Delivery of functional nanoparticles such as conjugation with quantum dots enables real-time fluorescence tracking and imaging. 8,9 Current delivery approaches are versatile, and each has it own unique advantages and limitations. For example, viralbased approaches can provide high transfer efficiency but are restricted to kilobase-sized nucleic acids with potential immunologic concerns.…”