Authors:

Sreesha Vedavalli Kidambi

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About Paper:

Delivering miRNA therapeutics to the brain poses a challenge due to the low permeability of the Blood-Brain Barrier (BBB). Many drugs are unable to achieve sufficient concentration in the brain to mitigate the symptoms of Alzheimer's Disease. Tetrahedral DNA nanostructures (TDN) are drug delivery vehicles with high potential to facilitate the transport of these therapeutics across the BBB. However, TDN are not easily uptaken by BBB cells because of DNA's hydrophilicity and the negative charge of DNA's phosphate backbone. Cationic lipids and polymers have been shown to mitigate these properties of DNA nanostructures and plasmid DNA. Because of this, functionalizing TDN with cationic lipids or polymers such as DOTAP, DOTMA, or DEAE- Dextran may allow for easier uptake of TDN into cells. To begin, DNA strands functionalized with Cy3 dye were assembled into TDN and characterized using gel electrophoresis. TDN were incubated with either DOTAP, DOTMA, DEAE-Dextran. Lipofectamine3000 was used as a positive control. Functionalized TDN (fTDN) were then tested for uptake efficiency against naked TDN in human kidney HEK293 as well as breast cancer CA1A cells, which are a field-standard positive control. Confocal microscopy was used to observe the uptake of fTDN into cells. This research addresses the difficulty of delivering therapeutics across the BBB by using TDN and testing the cellular uptake of fTDN. Further improvements include finding optimal concentrations of DOTAP, DOTMA, and DEAE-Dextran, incubating TDN with miRNA before functionalization to observe the uptake of miRNA into cells, and using BBB cells to test uptake of fTDN. Keywords: Drug Delivery; DNA Nanostructures

Source:

Purdue University / 2025

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Co-authors:

Sreesha Vedavalli Kidambi