Non-leaky vesicle fusion and enhanced cell transfection using a cationic facial amphiphile [19]

Journal of the American Chemical Society

Volumn 122, Issue 13, 2000, Pages 3252-3253

  Vandenburg, Yvonne R ^a   Smith, Bradley D ^a   Pérez Payán, M Nieves ^b   Davis, Anthony P ^b  

^a UNIVERSITY OF NOTRE DAME   (United States)

^b TRINITY COLLEGE DUBLIN   (Ireland)

Author keywords

[No Author keywords available]

Indexed keywords

AMPHOPHILE;

GENETIC TRANSFECTION; LETTER; LIPOPHILICITY; MEMBRANE FUSION; MEMBRANE VESICLE; PEPTIDE ANALYSIS; PH;

EID: 0034607456     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja994486z     Document Type: Letter

References (22)

1. Zimmerburg, J.; Chernomordik, L. V. Adv. Drug Delivery Rev. 1999, 38, 197-205.
2. Pécheur, E.-I.; Sainte-Marie, J.; Bienvenüe, A.; Hoekstra, D. J. Membr. Biol. 1999, 167, 1-17.
3. Fujii, G. Adv. Drug Delivery Rev. 1999, 38, 257-277.
4. Davies, S. M. A.; Kelly, S. M.; Price, N. C.; Bradshaw, J. P. FEBS Lett. 1998, 425, 415-418.
5. For another example that uses a different architecture, see: Zhang, Z. Y.; Smith, B. D., manuscript submitted.
6. Broderick, S.; Davis, A. P.; Williams, R. P. Tetrahedron Lett. 1998, 39, 6083-6086.
7. (a) Venkatesan, P.; Cheng, Y.; Kahne, D. J. Am. Chem. Soc. 1994, 116, 6955-6956.
8. (b) McQuade, D. T.; Barrett, D. G.; Desper, J. M.; Hayashi, R. K.; Gellman, S. H. J. Am. Chem. Soc. 1995, 117, 4862-4869.
9. (c) Li, C.; Peters, A. S.; Meredith, E. L.; Altman, G. W.; Savage, P. B. J. Am. Chem. Soc. 1998, 120, 2961-2962.
10. The synthesis of 1 has been reported before (Davis, A. P.; Pérez-Payán, M. N. Synlett 1999, S1, 991-993). The syntheses of 2 and 3 are described in the Supporting Information.
11. Abbreviations used in this report: ANTS, 1-aminonaphthalene-3,6,8-trisulfonate; DOPE, dioleoylphosphatidylethanolamine; DOTMA, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride; DPX, N,N′-p-xylenebis-(pyridinium bromide); NBD-PE, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitrobenz-2-oxa-1,3- diazol-4-yl); PC, egg phosphatidylcholine; PE, egg phosphalidylethanolamine; POPA, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidic acid; Rh-PE, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl); TES, N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid.
12. The modified literature methods (Düzgünes, N.; Wilschut, J. Methods Enzymol. 1993, 220, 3-14) used to measure lipid mixing, aqueous contents intermixing, and leakage are described in the Supporting Information. In short, lipid mixing was monitored by the probe dilution assay which uses the fluorescently labeled phospholipid, NBD-PE, and its resonance energy transfer quencher, Rh-PE. One vesicle population containing 0.3% of each the probes, NBD-PE and Rh-PE, is added to another population that is unlabeled. Lipid mixing is indicated by an increase in NBD-PE fluorescence intensity due to diminished quenching as the two probes are diluted. The contents mixing assay starts with two populations of vesicles, one encapsulating the fluorophore, ANTS, and the other containing the collisional quencher DPX. Fusion and mixing of aqueous contents results in a decrease in ANTS fluorescence intensity. The leakage assay starts with vesicles encapsulating a mixture of ANTS and DPX. Leakage of aqueous contents results in an increase in ANTS fluorescence intensity. All assays were reproduced independently at least once with an average uncertainty of less than 10%.
13. (a) Pécheur, E.-I.; Hoekstra, D.; Sainte-Marie, J.; Martin, I.; Bienvenüe, A.; Philippot, J. R. Biochemistry 1997, 36, 3773-3781.
14. (b) Pécheur, E.-I.; Martin, I.; Ruysschaert, J.-M.; Bienvenüe, A.; Hoekstra, D. Biochemistry 1998, 37, 2361-2371.
15. (c) Pécheur, E. I.; Sainte-Marie, J.; Bienvenüe, A.; Hoekstra, D. Biochemistry 1999, 38, 364-373.
16. (d) Martin, I.; Pécheur, E. I.; Ruysschaert, J. M.; Hoekstra, D. Biochemistry 1999, 38, 9337-9347.
17. Control studies show that there is no loss of amphiphiles 2 and 3 from the vesicles after dialysis for 12 h, and only 15% loss of amphiphile 1 (see Supporting Information). Thus, the difference in fusogenic properties is probably not due to any difference in amphiphile/vesicle affinity, but more likely to a difference in amphiphile/membrane orientation.
18. Fahey, D. A.; Carey, M. C.; Donovan, J. M. Biochemistry 1995, 34, 10886-10897.
19. Epand, R. M. Biochim. Biophys. Acta 1998, 1376, 353-368.
20. Miller, A. D. Angew. Chem., Int. Ed. 1998, 37, 1769-1785.
21. The literature contains a single report of successful transfection using a facial amphiphile, but the design (glycosylated cholate with polycationic tail) is quite different to the structure of 2. Walker, S.; Sofia, M. J.; Kakarla, R.; Kogan, N. A.; Wierichs, L.; Longley, C. B.; Bruker, K.; Axelrod, H. R.; Midha, S.; Babu, S.; Kahne, D. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 1585-1590.
22. 9 is a well-known transfection adjuvant and is often included in lipofection formulations. Hope, M. J.; Mui, B.; Ansell, S.; Ahkong, Q. F. Mol. Membr. Biol. 1998, 15, 1-14.