Description
- Abstract:
- In recent years, molecular beacons have drawn much attention from researchers due to their ability to image real time cellular mRNA expression. Molecular beacons are short hairpin oligonucleotides that fluoresce when bound to the mRNA of a gene of interest. Despite the utility of molecular beacons, current methods of cellular delivery lead to fluorescent debris that complicates analysis. This study investigates whether polymer-based delivery methods can improve upon current beacon delivery. We aimed to investigate whether two of these proposed methods, a diblock copolymer carrier system and a p(FA:SA) nanoencapsulation method, successfully deliver molecular beacons to multiple cell types while retaining functionality upon intracellular release. Beacons were delivered using a polymer developed for siRNA delivery, diblock copolymer. HEK293T and MG-63 cells were fixed and imaged after treatment with a beacon-polymer complex. A custom MATLAB script was used to quantify delivery by associating DAPI-stained nuclei with beacon signal. The number of cells expressing beacon fluorescence increased over a 24 hour period for all beacon sequences delivered with diblock copolymer. At 24 hours, beacon conditions all displayed fluorescent cells while beacon-only, diblock copolymer-only, and no treatment conditions exhibited none. Another polymer originally developed for drug delivery, p(FA:SA), was used to encapsulate beacons in preparation for functionality analysis. p(FA:SA) cellular treatment resulted in minimal cytotoxicity, while post-encapsulation analysis indicated an unquantifiable, low loading efficiency. Though further research is required to verify the efficacy of these two delivery methods, if successful, they can enable the study a myriad of cellular processes using molecular beacons.
- Notes:
- Concentration: Biology (ScB)/Physiology and Biotechnology
Access Conditions
- Rights
- In Copyright
- Restrictions on Use
- Collection is open for research.
Citation
Holman, Corey,
"Molecular beacon delivery methods in 2D cell culture"
(2016).
Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations, Biology and Medicine Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://doi.org/10.7301/Z0J101CQ
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Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations
Theses and Dissertations for the Molecular Pharmacology, Physiology, and Biotechnology department.... -
Biology and Medicine Theses and Dissertations
Theses and Dissertations for the Biology and Medicine department....