A method for quantifying cellular uptake of fluorescently labeled siRNA

Journal of Controlled Release

Volumn 148, Issue 1, 2010, Pages 106-109

  Vader, Pieter ^a   van der Aa, Leonardus J ^b   Engbersen, Johan F J ^b   Storm, Gert ^a   Schiffelers, Raymond M ^a  

^a UTRECHT UNIVERSITY   (Netherlands)

^b UNIVERSITY OF TWENTE   (Netherlands)

Author keywords

Fluorescence; Lipoplex; Polyplex; Quantitative uptake; Quenching; SiRNA

Indexed keywords

BIOLOGICAL STUDIES; CELLULAR UPTAKE; DELIVERY SYSTEMS; FLUORESCENT SIGNALS; INTRACELLULAR DELIVERY; LIPOPLEXES; LYSIS BUFFER; POLYPLEXES; QUANTITATIVE DETERMINATIONS; QUANTITATIVE UPTAKE; QUENCHING PHENOMENA; SIMPLE APPROACH; SIRNA; TRANSFECTED CELLS;

COMPLEXATION; FLUORESCENCE; QUENCHING;

RNA;

LIPOPLEX; SMALL INTERFERING RNA;

ARTICLE; CELL LYSATE; CYTOLYSIS; DRUG DELIVERY SYSTEM; DRUG FORMULATION; FLOW CYTOMETRY; FLUORESCENCE MICROSCOPY; PRIORITY JOURNAL;

CELL LINE, TUMOR; FLUORESCENT DYES; GENE TRANSFER TECHNIQUES; HUMANS; MICROSCOPY, FLUORESCENCE; RNA, SMALL INTERFERING;

EID: 78649529300     PISSN: 01683659     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.jconrel.2010.06.019     Document Type: Article

References (21)

1. Fire A., Xu S., Montgomery M.K., Kostas S.A., Driver S.E., Mello C.C. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998, 391:806-811.
2. Elbashir S.M., Harborth J., Lendeckel W., Yalcin A., Weber K., Tuschl T. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 2001, 411:494-498.
3. de Fougerolles A., Vornlocher H.P., Maraganore J., Lieberman J. Interfering with disease: a progress report on siRNA-based therapeutics. Nat. Rev. Drug Discov. 2007, 6:443-453.
4. Schiffelers R.M., Woodle M.C., Scaria P. Pharmaceutical prospects for RNA interference. Pharm. Res. 2004, 21:1-7.
5. Aigner A. Delivery systems for the direct application of siRNAs to induce RNA interference (RNAi) in vivo. J. Biomed. Biotechnol. 2006, 15.
6. Overhoff M., Wunsche W., Sczakiel G. Quantitative detection of siRNA and single-stranded oligonucleotides: relationship between uptake and biological activity of siRNA. Nucleic Acids Res. 2004, 32:e170.
7. Wolfrum C., Shi S., Jayaprakash K.N., Jayaraman M., Wang G., Pandey R.K., Rajeev K.G., Nakayama T., Charrise K., Ndungo E.M., Zimmermann T., Koteliansky V., Manoharan M., Stoffel M. Mechanisms and optimization of in vivo delivery of lipophilic siRNAs. Nat. Biotechnol. 2007, 25:1149-1157.
8. Liu W.L., Stevenson M., Seymour L.W., Fisher K.D. Quantification of siRNA using competitive qPCR. Nucleic Acids Res. 2009, 37:e4.
9. Kumar P., Ban H.S., Kim S.S., Wu H., Pearson T., Greiner D.L., Laouar A., Yao J., Haridas V., Habiro K., Yang Y.G., Jeong J.H., Lee K.Y., Kim Y.H., Kim S.W., Peipp M., Fey G.H., Manjunath N., Shultz L.D., Lee S.K., Shankar P. T cell-specific siRNA delivery suppresses HIV-1 infection in humanized mice. Cell 2008, 134:577-586.
10. Lin C., Zhong Z., Lok M.C., Jiang X., Hennink W.E., Feijen J., Engbersen J.F.J. Novel bioreducible poly(amido amine)s for highly efficient gene delivery. Bioconjug. Chem. 2007, 18:138-145.
11. Luten J., Akeroyd N., Funhoff A., Lok M.C., Talsma H., Hennink W.E. Methacrylamide polymers with hydrolysis-sensitive cationic side groups as degradable gene carriers. Bioconjug. Chem. 2006, 17:1077-1084.
12. Segura T., Hubbell J.A. Synthesis and in vitro characterization of an ABC triblock copolymer for siRNA delivery. Bioconjug. Chem. 2007, 18:736-745.
13. Breunig M., Hozsa C., Lungwitz U., Watanabe K., Umeda I., Kato H., Goepferich A. Mechanistic investigation of poly(ethylene imine)-based siRNA delivery: disulfide bonds boost intracellular release of the cargo. J. Control. Release 2008, 130:57-63.
14. Chen A.A., Derfus A.M., Khetani S.R., Bhatia S.N. Quantum dots to monitor RNAi delivery and improve gene silencing. Nucleic Acids Res. 2005, 33:e190.
15. Li S.D., Chono S., Huang L. Efficient gene silencing in metastatic tumor by siRNA formulated in surface-modified nanoparticles. J. Control. Release 2008, 126:77-84.
16. Merkel O.M., Beyerle A., Librizzi D., Pfestroff A., Behr T.M., Sproat B., Barth P.J., Kissel T. Nonviral siRNA delivery to the lung: investigation of PEG-PEI polyplexes and their in vivo performance. Mol. Pharm. 2009, 6:1246-1260.
17. Shim M.S., Kwon Y.J. Acid-responsive linear polyethylenimine for efficient specific, and biocompatible siRNA delivery. Bioconjug. Chem. 2009, 20:488-499.
18. Hoon Jeong J., Christensen L.V., Yockman J.W., Zhong Z., Engbersen J.F., Jong Kim W., Feijen J., Wan Kim S. Reducible poly(amido ethylenimine) directed to enhance RNA interference. Biomaterials 2007, 28:1912-1917.
19. Dunne J., Drescher B., Riehle H., Hadwiger P., Young B.D., Krauter J., Heidenreich O. The apparent uptake of fluorescently labeled siRNAs by electroporated cells depends on the fluorochrome. Oligonucleotides 2003, 13:375-380.
20. Chiu Y.L., Ali A., Chu C.Y., Cao H., Rana T.M. Visualizing a correlation between siRNA localization, cellular uptake, and RNAi in living cells. Chem. Biol. 2004, 11:1165-1175.
21. Lundberg P., El-Andaloussi S., Sutlu T., Johansson H., Langel U. Delivery of short interfering RNA using endosomolytic cell-penetrating peptides. FASEB J. 2007, 21:2664-2671.