Optimization of liposome mediated transfection of a neuronal cell line

被引:22
作者
McQuillin, A
Murray, KD
Etheridge, CJ
Stewart, L
Cooper, RG
Brett, PM
Miller, AD
Gurling, HMD
机构
[1] UCL, SCH MED,WINDEYER INST MED SCI, DEPT PSYCHIAT & BEHAV SCI,MOL PSYCHIAT LAB, LONDON W1P 6DB, ENGLAND
[2] UNIV LONDON IMPERIAL COLL SCI TECHNOL & MED, DEPT CHEM, LONDON SW7 2AY, ENGLAND
[3] ST THOMAS HOSP, SCH MED, RAYNE INST, LONDON SE1 7EH, ENGLAND
关键词
DC-Chol/DOPE; liposome; nerve; transfection;
D O I
10.1097/00001756-199704140-00031
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
A cell line derived from sensory neurons was transfected with high efficiency using cationic liposomes, formulated from 3 beta[N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol) and dioleoyl L-alpha-phosphatidylethanolamine (DOPE). This is the first time that cationic liposomes of this type have been reported to transfect a neuronal cell line. We used a reporter gene construct expressing beta-galactosidase under the control of the cytomegalovirus immediate early promoter and routinely observed transfection efficiencies > 40%. Parameters affecting transfection efficiency were examined and the ratio of DNA to liposome proved to be crucial. Liposome formulation procedures and cell transfection protocols devised here will be used as a basis for further in vivo and in vitro work.
引用
收藏
页码:1481 / 1484
页数:4
相关论文
共 12 条
[1]   NONINVASIVE LIPOSOME-MEDIATED GENE DELIVERY CAN CORRECT THE ION-TRANSPORT DEFECT IN CYSTIC-FIBROSIS MUTANT MICE [J].
ALTON, EWFW ;
MIDDLETON, PG ;
CAPLEN, NJ ;
SMITH, SN ;
STEEL, DM ;
MUNKONGE, FM ;
JEFFERY, PK ;
GEDDES, DM ;
HART, SL ;
WILLIAMSON, R ;
FASOLD, KI ;
MILLER, AD ;
DICKINSON, P ;
STEVENSON, BJ ;
MCLACHLAN, G ;
DORIN, JR ;
PORTEOUS, DJ .
NATURE GENETICS, 1993, 5 (02) :135-142
[2]  
CAPLEN NJ, 1995, GENE THER, V2, P603
[3]   LIPOSOME-MEDIATED CFTR GENE-TRANSFER TO THE NASAL EPITHELIUM OF PATIENTS WITH CYSTIC-FIBROSIS [J].
CAPLEN, NJ ;
ALTON, EWFW ;
MIDDLETON, PG ;
DORIN, JR ;
STEVENSON, BJ ;
GAO, X ;
DURHAM, SR ;
JEFFERY, PK ;
HODSON, ME ;
COUTELLE, C ;
HUANG, L ;
PORTEOUS, DJ ;
WILLIAMSON, R ;
GEDDES, DM .
NATURE MEDICINE, 1995, 1 (01) :39-46
[4]   THE ROLE OF DIOLEOYL PHOSPHATIDYLETHANOLAMINE IN CATIONIC LIPOSOME-MEDIATED GENE-TRANSFER [J].
FARHOOD, H ;
SERBINA, N ;
HUANG, L .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1995, 1235 (02) :289-295
[5]   Improvements in cationic liposomes for in vivo gene transfer [J].
Felgner, PL .
HUMAN GENE THERAPY, 1996, 7 (15) :1791-1793
[6]  
Gao X, 1995, GENE THER, V2, P710
[7]   A NOVEL CATIONIC LIPOSOME REAGENT FOR EFFICIENT TRANSFECTION OF MAMMALIAN-CELLS [J].
GAO, X ;
HUANG, L .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 179 (01) :280-285
[8]   DIRECT GENE-TRANSFER WITH DNA LIPOSOME COMPLEXES IN MELANOMA - EXPRESSION, BIOLOGIC ACTIVITY, AND LACK OF TOXICITY IN HUMANS [J].
NABEL, GJ ;
NABEL, EG ;
YANG, ZY ;
FOX, BA ;
PLAUTZ, GE ;
GAO, X ;
HUANG, L ;
SHU, SY ;
GORDON, D ;
CHANG, AE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (23) :11307-11311
[9]  
Sambrook J., 2002, MOL CLONING LAB MANU
[10]   Lipospermine-based gene transfer into the newborn mouse brain is optimized by a low lipospermine DNA charge ratio [J].
Schwartz, B ;
Benoist, C ;
Abdallah, B ;
Scherman, D ;
Behr, JP ;
Demeneix, BA .
HUMAN GENE THERAPY, 1995, 6 (12) :1515-1524