A Rapid Pathway Toward a Superb Gene Delivery System: Programming Structural and Functional Diversity into a Supramolecular Nanoparticle Library

Abstract

Nanoparticles are regarded as promising transfection reagents for effective and safe delivery of nucleic acids into specific type of cells or tissues providing an alternative manipulation/therapy strategy to viral gene delivery. However, the current process of searching novel delivery materials is limited due to conventional low-throughput and time-consuming multistep synthetic approaches. Additionally, conventional approaches are frequently accompanied with unpredictability and continual optimization refinements, impeding flexible generation of material diversity creating a major obstacle to achieving high transfection performance. Here we have demonstrated a rapid developmental pathway toward highly efficient gene delivery systems by leveraging the powers of a supramolecular synthetic approach and a custom-designed digital microreactor. Using the digital microreactor, broad structural/functional diversity can be programmed into a library of DNA-encapsulated supramolecular nanoparticles (DNA⊂SNPs) by systematically altering the mixing ratios of molecular building blocks and a DNA plasmid. In vitro transfection studies with DNA⊂SNPs library identified the DNA⊂SNPs with the highest gene transfection efficiency, which can be attributed to cooperative effects of structures and surface chemistry of DNA⊂SNPs. We envision such a rapid developmental pathway can be adopted for generating nanoparticle-based vectors for delivery of a variety of loads.

Light-Driven Droplet Manipulation Technologies for Lab-on-a-Chip Applications

“…Microfluidic devices promise a broad range of biomedical and chemical applications due to their potentials of small volume requirement, short analysis and diagnostic time, high sensitivity, and high throughput analysis [1][2][3]. Recently, particular attention has been paid to droplet-based (digital) microfluidic devices since droplets isolated in immiscible oil or air can be free from cross-contamination and dispersion [4].…”

Droplet Microfluidic Technique for the Study of Fermentation

“…Earlier, Song et al [17], Damean et al [18], Niu et al [19], Sun et al [20], Wang et al [21] and Agresti et al [22] reported systems to generate sequences of droplets scanning concentrations of one substance (up to four orders of magnitude) and used them to screening of biochemical reactions. Miller et al [23] demonstrated a microfluidic system that exploits Taylor-Aris dispersion to create a concentration gradient within about 10,000 droplets per compound.…”

A High‐Throughput Platform for Formulating and Screening Multifunctional Nanoparticles Capable of Simultaneous Delivery of Genes and Transcription Factors

“…Combining integrated microfluidic technologies [16][17][18][19][20][21][22][23][24][25] with a supramolecular synthetic strategy, [15,[26][27][28][29][30] we demonstrate herein a high-throughput approach for formulating and screening multifunctional supramolecular nanoparticles (MFSNPs) that are capable of simultaneous delivery of gene and functional proteins with superb efficiency and controllable stoichiometry among individual payloads ( Figure 1). As illustrated in Figure 1a, this approach utilized two microfluidic systems including a digital droplet generator (DDG; Supporting Information, Section 1.2-1.5, video available in Supporting Information) [20,22] and a microfluidic cell culture array [31,32] .…”

“…As illustrated in Figure 1a, this approach utilized two microfluidic systems including a digital droplet generator (DDG; Supporting Information, Section 1.2-1.5, video available in Supporting Information) [20,22] and a microfluidic cell culture array [31,32] . To enable the capability of on-chip synthesis of MFSNPs using the DDG, we developed a modular assembly system based on the supramolecular synthetic strategy [15,[26][27][28][29][30] (Figure 1b).…”

“…As illustrated in Figure 1b, the optimization studies using the model system consist of the following modules: (i) the protein modules include two different protein nanocapsules [5] (i.e., Cy5 labeled bovine serum albumin, BSA-Cy5, and rhodamin B-labeled horseradish peroxidase, HRP-RhB; Supporting Information, Section 1.8), which were made by encapsulating the protein within a thin layer of degradable polymer of which the surface was linked with Ad groups; (ii) the gene module can be any DNA of interest (i.e., EGFP-encode DNA plasmid, pEGFP); (iii) the function modules are a series of poly(ethylene glycol) (PEG) derivatives, [20,26,27] of which one endgroup is terminated with an Ad group and the other end is terminated with a methyl group (Ad-PEG [26] ; Supporting Information, Section 1.9) or a functional peptide, such as RGD (Ad-PEG-RGD [34] ; Supporting Information, Section 1.10) and TAT (Ad-PEG-TAT [35] ; Supporting Information, Section 1.11); and (iv) the scaffold module is CD-linked poly(ethylenimine) (CD-PEI [26] ; Supporting Information, Section 1.12). This strategy yields generally applicable protein modules whose assembly properties are not affected by the core protein.…”