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Cell Assay Protocol 001: Flow Cytometry to Test the Binding of RNA Nanoparticles with Cells

1. Culture cells in their complete medium. Dissociate the cells with trypsin and wash with PBS once.
2. Incubate 0.2 million cells with fluorescent labeled RNA nanoparticles at a final concentration of 50 nM to 200 nM in blank medium, at 37 °C for 1 to 4 hrs.
3. Wash cells with PBS, re-suspend cells in PBS buffer.
4. Assay the binding of RNA nanoparticles with cells by Flow Cytometry, decide excitation and mission spectrum based on the information from your product.
 
Reference:
1. Shu Y, Shu D, Haque F, Guo P. Fabrication of pRNA Nanoparticles to Deliver Therapeutic RNAs and Bioactive Compounds into Tumor Cells. Nature Protocols 2013;8:1635-1659.
2. Shu D, Li H, Shu Y, Xiong G, Carson WE, Haque F, Xu R, Guo P. Systemic Delivery of Anti-miRNA for Suppression of Triple Negative Breast Cancer Utilizing RNA Nanotechnology. ACS Nano 2015; 9:9731-9740.
3. Shu D, Shu Y, Haque F, Abdelmawla S, Guo P. Thermodynamically stable RNA three-way junction for constructing multifunctional nanoparticles for delivery of therapeutics. Nature Nanotechnology 2011; 6:658-67.
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