Current cancer research has begun to focus on analyzing micro-RNA (miRNA) as biological markers for cancerous cells and their potential as diagnostic markers. Specific miRNA sequences can be identified as targets for DNA-based fluorescent sensors (molecular beacons). These sensors can be introduced into cell cultures using chemical transfection, which while well-established it has not been confirmed whether DNA-duplexes are transfected as individual strands or hybridized. A molecular beacon miR-21 (an oncogenic miRNA) was tested at each stage of the transfection protocol using Lipofectamine MessengerMAX (MM) to determine the potential for strand dissociation under transfection conditions by fluorescence spectroscopy. Initial work in sodium phosphate buffer found no significant strand dissociation upon the addition of MM. In cell culture media, however, there was increased strand dissociation in both un-supplemented DMEM and in DMEM supplemented with FBS and antibiotics. This preliminary data indicates that the transfection reagent MM may cause dissociation on complex formation, prior to entry into the cell. Identifying the state of DNA during transfection could prove crucial in the use of multi-stranded fluorescent sensing systems.

Effects of liopfectamine MessengerMAX on hybridization state of DNA-based molecular beacons

255th National Meeting (2018)

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