Can we better understand the intracellular behavior of DNA nanoparticles by fluorescence correlation spectroscopy?
The use of non-viral gene carriers to deliver small nucleic acids like antisense oligonucleotides (ODNs) and small interfering RNA (siRNA) remains an attractive but challenging goal in antisense therapy. Indeed, different barriers need to be overcome in the delivery process before a therapeutic effect can be obtained. One promising technique which we have been evaluating to improve our understanding of the intracellular behavior of nucleic acids/carrier complexes is Fluorescence Correlation Spectroscopy (FCS). In particular, we have used FCS for studying the protection of the nucleic acids against enzymatic degradation, and the association and dissociation of the nucleic acids with their carrier, both in buffer and in living cells. In this report, we will review our experience and findings on the use of FCS for that purpose and discuss the strengths and weaknesses of this interesting technique.
Remaut
K
K
Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium.
Lucas
B
B
Raemdonck
K
K
Braeckmans
K
K
Demeester
J
J
De Smedt
S C
SC
D000818:Animals; D002021:Buffers; D002232:Carbocyanines / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D016264:Dextran Sulfate / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008956:Models, Chemical; D053758:Nanoparticles / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000494:pharmacology; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D014709:Vero Cells
D002021:Buffers; D002232:Carbocyanines; D005456:Fluorescent Dyes; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D034741:RNA, Small Interfering; C085321:cyanine dye 5; D004247:DNA; D016264:Dextran Sulfate; D012259:Ribonuclease, Pancreatic; D016650:Fluorescein-5-isothiocyanate