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Liposomes pp 425–437Cite as

Fluorescence Methods for Evaluating Lipoplex-Mediated Gene Delivery

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 606))

Abstract

The biological activity of cationic liposome/DNA complexes (“lipoplexes”) is strongly dependent on their ability to protect DNA and to interact with cells, including binding to the cell surface, internalization via endocytosis and cytoplasmic delivery of the DNA. In this chapter, we describe a number of methods and procedures to study these processes, based on the use of fluorescent probes.

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References

  1. Wasungu L, Hoekstra D (2006) Cationic lipids, lipoplexes and intracellular delivery of genes. J Control Release 116:255-264

    Article  CAS  PubMed  Google Scholar 

  2. Khalil IA, Kogure K, Akita H, Harashima H (2006) Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery. Pharmacol Rev 58:32-45

    Article  CAS  PubMed  Google Scholar 

  3. Pedroso de Lima MC, Simões S, Pires P, Faneca H, Düzgünes˛ N (2001) Cationic lipid-DNA complexes in gene delivery: from biophysics to biological applications. Adv Drug Deliv Rev 47:277-294

    Article  CAS  PubMed  Google Scholar 

  4. Simões S, Filipe A, Faneca H, Mano M, Penacho N, Düzgünes˛ N et al (2005) Cationic liposomes for gene delivery. Expert Opin Drug Deliv 2:237-254

    Article  PubMed  Google Scholar 

  5. Lechardeur D, Lukacs GL (2002) Intracellular barriers to non-viral gene transfer. Curr Gene Ther 2:183-194

    Article  CAS  PubMed  Google Scholar 

  6. Faneca H, Cabrita AS, Simões S, Pedroso de Lima MC (2007) Evaluation of the antitumoral effect mediated by IL-12 and HSV-tk genes when delivered by a novel lipid-based system. Biochim Biophys Acta 1768:1093-1102

    Article  CAS  PubMed  Google Scholar 

  7. Faneca H, Simões S, Pedroso de Lima MC (2004) Association of albumin or protamine to lipoplexes: enhancement of transfection and resistance to serum. J Gene Med 6:681-692

    Article  CAS  PubMed  Google Scholar 

  8. Simões S, Slepushkin V, Pires P, Gaspar R, Pedroso de Lima MC, Düzgünes˛ N (2000) Human serum albumin enhances DNA transfection by lipoplexes and confers resistance to inhibition by serum. Biochim Biophys Acta 1463:459-469

    Article  PubMed  Google Scholar 

  9. Wang L, MacDonald RC (2004) Effects of microtubule-depolymerizing agents on the transfection of cultured vascular smooth muscle cells: Enhanced expression with free drug and especially with drug-gene lipoplexes. Mol Ther 9:729-737

    Article  CAS  PubMed  Google Scholar 

  10. Hasegawa S, Hirashima N, Nakanishi M (2001) Microtubule involvement in the intracellular dynamics for gene transfection mediated by cationic liposomes. Gene Ther 8:1669-1673

    Article  CAS  PubMed  Google Scholar 

  11. Chowdhury NR, Hays RM, Bommineni VR, Franki N, Chowdhury JR, Wu CH et al (1996) Microtubular disruption prolongs the expression of human bilirubin-uridinediphos­phoglucuronate-glucuronosyltransferase-1 gene transferred into Gunn rat livers. J Biol Chem 271:2341-2346

    Article  CAS  PubMed  Google Scholar 

  12. Faneca H, Simões S, Pedroso de Lima MC (2002) Evaluation of lipid-based reagents to mediate intracellular gene delivery. Biochim Biophys Acta 1567:23-33

    Article  CAS  PubMed  Google Scholar 

  13. Faneca H, Faustino A, Pedroso de Lima MC (2008) Synergistic antitumoral effect of non-viral HSV-tk/GCV gene therapy and vinblastine in mammary adenocarcinoma cells. J Control Release 126:175-184

    Article  CAS  PubMed  Google Scholar 

  14. Pedroso de Lima MC, Faneca H, Mano M, Penacho N, Düzgünes˛ N, Simões S (2003) Biophysical characterization of cationic liposome-DNA complexes and their interaction with cells. Meth Enzymol 373:298-312

    Article  CAS  Google Scholar 

  15. Fiske CH, Subbarow Y (1925) The calori­metric determination of phosphorus. J Biol Chem 66:375-400

    Google Scholar 

  16. Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466-468

    CAS  PubMed  Google Scholar 

  17. Düzgünes˛ N (2003) Preparation and quantitation of small unilamellar liposomes and large unilamellar reverse-phase evaporation liposomes. Meth Enzymol 367:23-27

    Article  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Faculty of Science and Technology, Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal

    Henrique Faneca

  2. Department of Microbiology, Arthur A. Dugoni School of Dentistry, University of the Pacific, San Francisco, CA, USA

    Nejat Düzgüneş

  3. Department of Biochemistry, Faculty of Science and Technology, Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal

    Maria C. Pedroso de Lima

Authors
  1. Henrique Faneca
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  2. Nejat Düzgüneş
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  3. Maria C. Pedroso de Lima
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Editor information

Editors and Affiliations

  1. College of Pharmacy, Midwestern University, N. 59th Avenue 19555, Glendale, 85308, U.S.A.

    Volkmar Weissig

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Faneca, H., Düzgüneş, N., de Lima, M.C.P. (2010). Fluorescence Methods for Evaluating Lipoplex-Mediated Gene Delivery. In: Weissig, V. (eds) Liposomes. Methods in Molecular Biology™, vol 606. Humana Press. https://doi.org/10.1007/978-1-60761-447-0_29

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  • DOI: https://doi.org/10.1007/978-1-60761-447-0_29

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-446-3

  • Online ISBN: 978-1-60761-447-0

  • eBook Packages: Springer Protocols

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