A Multi‐RNAi Microsponge Platform for Simultaneous Controlled Delivery of Multiple Small Interfering RNAs

Abstract

Packaging multiple small interfering RNA (siRNA) molecules into nanostructures at precisely defined ratios is a powerful delivery strategy for effective RNA interference (RNAi) therapy. We present a novel RNA nanotechnology based approach to produce multiple components of polymerized siRNA molecules that are simultaneously self‐assembled and densely packaged into composite sponge‐like porous microstructures (Multi‐RNAi‐MSs) by rolling circle transcription. The Multi‐RNAi‐MSs were designed to contain a combination of multiple polymeric siRNA molecules with precisely controlled stoichiometry within a singular microstructure by manipulating the types and ratios of the circular DNA templates. The Multi‐RNAi‐MSs were converted into nanosized complexes by polyelectrolyte condensation to manipulate their physicochemical properties (size, shape, and surface charge) for favorable delivery, while maintaining the multifunctional properties of the siRNAs for combined therapeutic effects. These Multi‐RNAi‐MS systems have great potential in RNAi‐mediated biomedical applications, for example, for the treatment of cancer, genetic disorders, and viral infections.

Nanoscale Self-Assembly for Therapeutic Delivery

“…So these DNA motifs have been used to trigger host immune response as these are recognized by TLR-9 located on endosomes of host membrane of immune system which activates the innate immune pathway of host immune system. Similarly, siRNA and miRNA delivery have also been carried out using DNA nanostructures for gene silencing applications (Lee et al, 2012;Fakhoury et al, 2013;Bujold et al, 2016;Roh et al, 2016;Qian et al, 2017;Nahar et al, 2018). Various types of DNA nanostructures, that have been used as delivery vehicles, are listed in Table 4 (Hu et al, 2018;Madhanagopal et al, 2018).…”

Nucleic Acid–Based Functional Nanomaterials as Advanced Cancer Therapeutics

“…Similar to RCA, RCT produced long single‐strand RNA with preprogrammed circular DNA template in the presence of RNA polymerases through rolling circle amplification . As an efficient enzymatic reaction to generate multiple as well as repetitive designed RNA sequences, RCT has also applied to the field of siRNA delivery …”

Visualization of the intracellular location and stability of DNA flowers with a label-free fluorescent probe

“…Importantly, DNA ower constructs were observed to enter RAW264.7 cells and accumulate in lysosomes, which is signicant to rationally design nanocarriers fated to the macrophages and the lysosomes, such as immunostimulatory moieties delivery vehicles 8 or nanovaccines. 9…”

“…Generating a composite nucleic acid/magnesium pyrophosphate ower-like structure and organizing functional materials, RCA or RCT-derived constructs have been fabricated for a variety of biological applications, such as biological assays, 3 enhanced biocatalytic activity, 4 multiplexed cellular imaging 5 and traceable targeted drug delivery, 6 antisense oligonucleotide delivery 7 and siRNA delivery 8,9 for cancer therapy, antigen and adjuvant delivery 10,11 for cancer immunotherapy. Along with the utility of these ower-like materials, it is necessary to further investigate the interactions between RCA/RCT-derived structures and live cells.…”