Sugar-based tertiary amino gemini surfactants with a vesicle-to-micelle transition in the endosomal pH range mediate efficient transfection in vitro

被引:123
作者
Fielden, ML
Perrin, C
Kremer, A
Bergsma, M
Stuart, MC
Camilleri, P
Engberts, JBFN [1 ]
机构
[1] Univ Groningen, Stratingh Inst, Phys Organ Chem Unit, NL-9747 AG Groningen, Netherlands
[2] SmithKline Beecham Pharmaceut, Harlow CM19 5AD, Essex, England
[3] Univ Groningen, Dept Biophys Chem, NL-9700 AB Groningen, Netherlands
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 2001年 / 268卷 / 05期
关键词
amphiphile; carbohydrate; transfection; vesicle-to-micelle transition;
D O I
10.1046/j.1432-1327.2001.01995.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Novel reduced sugar gemini amphiphiles linked through their tertiary amino head groups via alkyl spacers of 4 or 6 carbons, and with varying (unsaturated) alkyl tail lengths of 12-18, have been synthesized and tested for transfection in vitro in an adherent Chinese hamster ovary cell line (CHO-K1). Transfection efficiencies peaked at 2.7 times that of the commercial standard Lipofectamine Plus/2000 for pure solutions of the compound bearing unsaturated (oleyl) alkyl tails. For those compounds bearing saturated alkyl tails, transfection efficiency peaked at a tail length of 16, at a level similar to Lipofectamine Plus/2000. All of the amphiphiles formed bilayer vesicles at physiological pH. Some of the amino groups at the surface were protonated, and vesicles therefore bore a positive charge. Increased protonation with reduced pH resulted in greatly increased monomer solubility and a morphology change from vesicle to micelle at characteristic pH values, dependent on the tail length. For the compounds promoting high transfection efficiency, this characteristic pH was within the range found in the endosomal compartment (7.4-4.0). Formation of mixed micelles between gemini surfactant and membrane phospholipids at reduced pH may therefore provide a method of endosome rupture and subsequent escape of entrapped DNA, thus discarding the need for extra fusogenic or endosomolytic agents. The positive charge on the vesicles at physiological pH drives the colloidal association with DNA. Small angle X-ray scattering measurements indicate that lamellar aggregates are formed, which have a d spacing of 48-54 Angstrom. Preliminary differential scanning calorimetric measurements suggest that reduction of pH causes a disordering of the hydrocarbon region of the DNA-surfactant complex.
引用
收藏
页码:1269 / 1279
页数:11
相关论文
共 57 条
[31]   CLINICAL RESEARCH - LESS HYPE, MORE BIOLOGY NEEDED FOR GENE-THERAPY [J].
MARSHALL, E .
SCIENCE, 1995, 270 (5243) :1751-1751
[32]  
Meekel AAP, 2000, EUR J ORG CHEM, V2000, P665
[33]   Physico-chemical aspects of the interaction between DNA and oppositely charged mixed liposomes [J].
Mel'nikova, YS ;
Mel'nikov, SM ;
Löfroth, JE .
BIOPHYSICAL CHEMISTRY, 1999, 81 (02) :125-141
[34]   SPECIFIC GENE-TRANSFER MEDIATED BY LACTOSYLATED POLY-L-LYSINE INTO HEPATOMA-CELLS [J].
MIDOUX, P ;
MENDES, C ;
LEGRAND, A ;
RAIMOND, J ;
MAYER, R ;
MONSIGNY, M ;
ROCHE, AC .
NUCLEIC ACIDS RESEARCH, 1993, 21 (04) :871-878
[35]  
Miller AD, 1998, ANGEW CHEM INT EDIT, V37, P1769
[36]   HUMAN GENE-THERAPY COMES OF AGE [J].
MILLER, AD .
NATURE, 1992, 357 (6378) :455-460
[37]   STRUCTURE AND PHARMACOLOGY OF THE PROTON-ATPASES [J].
NELSON, N .
TRENDS IN PHARMACOLOGICAL SCIENCES, 1991, 12 (02) :71-75
[38]   ELECTROPORATION - APPLICATION TO HUMAN LYMPHOID-CELL LINES FOR STABLE INTRODUCTION OF A TRANSACTIVATOR GENE OF HUMAN T-CELL LEUKEMIA-VIRUS TYPE-I [J].
OHTANI, K ;
NAKAMURA, M ;
SAITO, S ;
NAGATA, K ;
SUGAMURA, K ;
HINUMA, Y .
NUCLEIC ACIDS RESEARCH, 1989, 17 (04) :1589-1604
[39]   Physico-chemical characterization of a double long-chain cationic amphiphile (Vectamidine) by microelectrophoresis [J].
Pector, V ;
Caspers, J ;
Banerjee, S ;
Deriemaeker, L ;
Fuks, R ;
El Ouahabi, A ;
Vandenbranden, M ;
Finsy, R ;
Ruysschaert, JM .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1998, 1372 (02) :339-346
[40]   Nonionic bolaamphiphiles and gemini surfactants based on carbohydrates [J].
Pestman, JM ;
Terpstra, KR ;
Stuart, MCA ;
vanDoren, HA ;
Brisson, A ;
Kellogg, RM ;
Engberts, JBFN .
LANGMUIR, 1997, 13 (25) :6857-6860