Molecular Recognition Enables Nanosubstrate-Mediated Delivery of Gene-Encapsulated Nanoparticles with High Efficiency

Abstract

Substrate-mediated gene delivery is a promising method due to its unique ability to preconcentrate exogenous genes onto designated substrates. However, many challenges remain to enable continuous and multiround delivery of the gene using the same substrates without depositing payloads and immobilizing cells in each round of delivery. Herein we introduce a gene delivery system, nanosubstrate-mediated delivery (NSMD) platform, based on two functional components with nanoscale features, including (1) DNA⊂SNPs, supramolecular nanoparticle (SNP) vectors for gene encapsulation, and (2) Ad-SiNWS, adamantane (Ad)-grafted silicon nanowire substrates. The multivalent molecular recognition between the Ad motifs on Ad-SiNWS and the β-cyclodextrin (CD) motifs on DNA⊂SNPs leads to dynamic assembly and local enrichment of DNA⊂SNPs from the surrounding medium onto Ad-SiNWS. Subsequently, once cells settled on the substrate, DNA⊂SNPs enriched on Ad-SiNWS were introduced through the cell membranes by intimate contact with individual nanowires on Ad-SiNWS, resulting in a highly efficient delivery of exogenous genes. Most importantly, sequential delivery of multiple batches of exogenous genes on the same batch cells settled on Ad-SiNWS was realized by sequential additions of the corresponding DNA⊂SNPs with equivalent efficiency. Moreover, using the NSMD platform in vivo, cells recruited on subcutaneously transplanted Ad-SiNWS were also efficiently transfected with exogenous genes loaded into SNPs, validating the in vivo feasibility of this system. We believe that this nanosubstrate-mediated delivery platform will provide a superior system for in vitro and in vivo gene delivery and can be further used for the encapsulation and delivery of other biomolecules.

mRNA-Producing Pseudo-nucleus System

“…This mechanism, the so‐called the central dogma, derives its importance from the fact that it can be applied to metabolism‐mimicking synthetic cell designs in the same context. To date, however, the creation of synthetic cells has consisted of simply trapping exogenous genomes into polymer complexes and studying synthetic metabolism mimicry …”

Physical Delivery of Macromolecules using High-Aspect Ratio Nanostructured Materials

“…49 The combination of cyclodextrin-grafted branched PEI, adamantane-grafted polyamidoamine, adamantine-grafted poly(ethylene glycol, PEG), and Si NWs achieved efficient delivery of DNA supramolecular nanoparticles in mice. 49 …”

Micro‐ and Nanotechnologies for Intracellular Delivery

“…Since 2007, the majority of the research has been on in vitro applications of nanoneedles/nanowires. In early 2014, Tseng and co‐workers reported an in vivo study of substrate‐mediated gene delivery, as shown in Figure . In their delivery system, two nanoscale features were included: 1) DNA‐SNPs (supramolecular nanoparticles) as a vector for gene encapsulation, and 2) adamantane (Ad)‐grafted silicon nanowire substrates.…”

“…The Ad/CD recognition system is also responsible for the supramolecular assembly of DNA‐SNPs from the three molecular building blocks (i.e., CD‐PEI, Ad‐PAMAM, and Ad‐PEG) and plasmid DNA. Reproduced with permission . Copyright 2014, American Chemical Society.…”