This document summarizes a study examining the cellular uptake of siRNA nanoparticles for drug delivery in whole blood and lymphocytes. The study introduces fluorescently tagged RNA nanoparticles to both whole blood samples and isolated lymphocytes from human donors. Flow cytometry is used to measure fluorescence changes indicating the relative uptake of nanoparticles by blood cells. Preliminary results show a marked difference in nanoparticle uptake depending on whether the samples are diluted or undiluted, which may affect apoptosis and cell morphology of lymphocytes and their interaction with nanoparticles. Further experimentation will examine which nanoparticles and cells have the most efficient uptake, the best lipid-like carrier, and mechanisms of cellular entry.

1. CellularUptake of siRNA NanoparticlesforDrugDeliveryinWhole BloodandLymphocytes
The newestfrontierof drugdeliveryanddisease treatmentisnanotechnologyutilizingtherapeutic
nucleicacids,whichare provingtobe fullycustomizable,programmable andcancarry multiple
functionalities.Nucleicacidbasednanoparticlesfunctionalizedwithmultiple shortinterference RNAs
(siRNAs),orothertherapeuticoligonucleotidesandformulatedwithlipid-like carriersforefficient
intracellulardeliverycanactas an active pharmaceutical ingredient.Due tothe route of administration,
it isimportantto studyhowthisnewtherapeutictechnologyinteractswithbloodandlymphocytesto
elucidate possibleundesirable side-effects.
Thisinvestigationexaminesthe cellularuptake bywholebloodandbylymphocyte isolationsfrom
humandonorsof several fluorescentlytaggedfunctional RNA nanoparticlesselectedfromthe
laboratorylibrary.Lymphocytesare isolatedfromeachwhole bloodsample andthe siRNA nanoparticles
formulatedwithpolycationiccarriersare introducedtoboththe whole bloodand lymphocyte isolation.
Fluorescence changesindicatingthe relative uptake of nano-formulationsbybloodcellsare measured
viaflowcytometryandfurtheranalyzed.Experimentationhasthusfarshowna markeddifferencein
nanoparticle uptake whenthe formulationisintroducedafterthe samplesare seriallydilutedas
opposedtointroductiontoundilutedhumanbloodsamples.Thismayindicate thatthe currentlyused
experimental protocolmayaffectapoptosisandcell morphologyof lymphocytesthuspromoting their
interactionwithnanoparticles.Furtherexperimentationaimstoexaminewhichconstructsandcells
have the most efficientcellularuptake bybloodcells,whichlipid-like carrierworksbestforuptake and
mechanismsforcellularentry.