TY - JOUR
T1 - Lectin enhancement of the lipofection efficiency in human lung carcinoma cells
AU - Yanagihara, Katsunori
AU - Cheng, Pi Wan
N1 - Funding Information:
The authors wish to thank the NIH NIDDK-Cystic Fibrosis Foundation, the National Institutes of Health (HL RO1 56190) and State of Nebraska Department of Health (LB506 99-12), State of Nebraska (Research Initiative Grant on Gene Therapy Program) for supporting the research.
PY - 1999/10/18
Y1 - 1999/10/18
N2 - Poor transfection efficiency of human lung carcinoma cells by lipofection begs further development of more efficient gene delivery strategies. The purpose of this study was to determine whether lectins can improve the lipofection efficiency in lung carcinoma cells. A549, Calu3, and H292 cells grown to 90% confluence were transfected for 18 h with a plasmid DNA containing a β-galactosidase reporter gene (pCMVlacZ) using lipofectin plus a lectin as the vector. Ten different lectins, which exhibit a wide range of carbohydrate-binding specificities, were examined for their abilities to enhance the efficiency of lipofection. The transfected cells were assessed for transfection efficiency by β-galactosidase activity (units/μg protein) and % blue cells following X-Gal stain. Lipofectin supplemented with Griffonia simplicifolia-I (GS-I) yields largest enhancement of the lipofection efficiency in A549 and Calu3 cells (5.3- and 28-fold, respectively). Maackia amurensis gives the largest enhancement (6.5-fold) of lipofection efficiency in H292 cells. The transfection efficiency correlates with the amounts of DNA delivered to the nucleus. Binding of FITC-labeled GS- I and the enhancement of the lipofection efficiency by GS-I were inhibited by α-methyl-D-galactopyranoside, indicating an α-galactoside-mediated gene transfer to lung carcinoma cells. We conclude that lectin-facilitated lipofection is an efficient gene delivery strategy. Employment of cell type- specific lectins may allow for efficient cell type-specific gene targeting.
AB - Poor transfection efficiency of human lung carcinoma cells by lipofection begs further development of more efficient gene delivery strategies. The purpose of this study was to determine whether lectins can improve the lipofection efficiency in lung carcinoma cells. A549, Calu3, and H292 cells grown to 90% confluence were transfected for 18 h with a plasmid DNA containing a β-galactosidase reporter gene (pCMVlacZ) using lipofectin plus a lectin as the vector. Ten different lectins, which exhibit a wide range of carbohydrate-binding specificities, were examined for their abilities to enhance the efficiency of lipofection. The transfected cells were assessed for transfection efficiency by β-galactosidase activity (units/μg protein) and % blue cells following X-Gal stain. Lipofectin supplemented with Griffonia simplicifolia-I (GS-I) yields largest enhancement of the lipofection efficiency in A549 and Calu3 cells (5.3- and 28-fold, respectively). Maackia amurensis gives the largest enhancement (6.5-fold) of lipofection efficiency in H292 cells. The transfection efficiency correlates with the amounts of DNA delivered to the nucleus. Binding of FITC-labeled GS- I and the enhancement of the lipofection efficiency by GS-I were inhibited by α-methyl-D-galactopyranoside, indicating an α-galactoside-mediated gene transfer to lung carcinoma cells. We conclude that lectin-facilitated lipofection is an efficient gene delivery strategy. Employment of cell type- specific lectins may allow for efficient cell type-specific gene targeting.
KW - Gene therapy
KW - Gene transfer
KW - Lectin
KW - Liposome
KW - Non-viral vector
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U2 - 10.1016/S0304-4165(99)00100-2
DO - 10.1016/S0304-4165(99)00100-2
M3 - Article
C2 - 10572922
AN - SCOPUS:0032883606
SN - 0304-4165
VL - 1472
SP - 25
EP - 33
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 1-2
ER -