Conditional Deoxyribozyme-Nanoparticle Conjugates for miRNA-Triggered Gene Regulation.
Jiahui ZhangRong MaAaron BlanchardJessica PetreeHanjoong JoKhalid SalaitaPublished in: ACS applied materials & interfaces (2020)
DNA-nanoparticle (NP) conjugates have been used to knockdown gene expression transiently and effectively, making them desirable tools for gene regulation therapy. Because DNA-NPs are constitutively active and are rapidly taken up by most cell types, they offer limited control in terms of tissue or cell type specificity. To take a step toward solving this issue, we incorporate toehold-mediated strand exchange, a versatile molecular programming modality, to switch the DNA-NPs from an inactive state to an active state in the presence of a specific RNA input. Because many transcripts are unique to cell subtype or disease state, this approach could one day lead to responsive nucleic acid therapeutics with enhanced specificity. As a proof of concept, we designed conditional deoxyribozyme-nanoparticles (conditional DzNPs) that knockdown tumor necrosis factor α (TNFα) mRNA upon miR-33 triggering. We demonstrate toehold-mediated strand exchange and restoration of TNFα DNAzyme activity in the presence of miR-33 trigger, with optimization of the preparation, configuration, and toehold length of conditional DzNPs. Our results indicate specific and strong ON/OFF response of conditional DzNPs to the miR-33 trigger in buffer. Furthermore, we demonstrate endogenous miR-33-triggered knockdown of TNFα mRNA in mouse macrophages, implying the potential of conditional gene regulation applications using these DzNPs.
Keyphrases
- nucleic acid
- cell proliferation
- long non coding rna
- rheumatoid arthritis
- long noncoding rna
- gene expression
- circulating tumor
- single molecule
- cell free
- single cell
- cancer therapy
- cell therapy
- dna methylation
- stem cells
- binding protein
- high resolution
- climate change
- structural basis
- iron oxide
- circulating tumor cells
- liquid chromatography
- label free
- human health
- molecularly imprinted