Atelocollagen-based gene transfer in cells allows high-throughput screening of gene functions

被引:69
作者
Honma, K
Ochiya, T
Nagahara, S
Sano, A
Yamamoto, H
Hirai, K
Aso, Y
Terada, M
机构
[1] Natl Canc Ctr, Res Inst, Chuo Ku, Tokyo 1040045, Japan
[2] Koken Biosci Inst, Kita Ku, Tokyo 1150051, Japan
[3] Sumitomo Pharmaceut Co Ltd, Formulat Res Labs, Ibaraki, Osaka 5670878, Japan
关键词
atelocollagen; gene transfer; array; plasmid DNA; adenovirus; antisense; high-throughput;
D O I
10.1006/bbrc.2001.6133
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
We have previously demonstrated that Atelocollagen, used clinically for wound healing, is a reliable safe carrier for gene delivery. To obtain phenotypic changes by gene expression of cDNA, we developed an efficient technique for high-throughput gene transfer and expression screening in mammalian cells in microarrays by precoating a microplate with an Atelocollagen complexed with cDNA to which cells are then seeded. The complexes with a nanoparticle form were efficiently transduced into cells without use of any additional transfection reagent, and they allowed for long-term gene expression without apparent chromosomal integration. The complex spotted onto the well of a microplate was stable for a long period and allowed the cells to transduce and express reporter genes in a dose-dependent manner. We also showed that the present method using Atelocollagen-based gene transfer is applicable to gene medicines such as antisense ODNs and adenovirus vectors. These results suggest that Atelocollagen may be appropriate for general use in high-throughput screening of large sets of gene medicines for functional analyses in mammalian cells. (C) 2001 Elsevier Science.
引用
收藏
页码:1075 / 1081
页数:7
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