The role of atelocollagen-based cell transfection array in high-throughput screening of gene functions and in drug discovery.
The human genome project has been completed, but the function of many genes is unknown. It is, therefore, necessary to elucidate the function of a large number of genes within a short time. To achieve this goal, materials are needed that condense or package DNA into nano-particles that can easily be taken up by cells and would allow DNA to be retained without degradation. Atelocollagen is a reliable carrier for gene delivery because it is considered safe and appropriate for practical use. We developed a basic technique for high-throughput gene transfer and expression screening by pre-coating a multi-well plate with an Atelocollagen/DNA complex in which cells are then seeded. Complexes with a nano-particle form were efficiently transduced into cells without the use of additional transfection reagents, and they allowed for long-term gene expression. The complex spotted onto the well of a plate was stable for a long period and allowed the cells to transduce and express reporter genes. We also showed that the present method with Atelocollagen-based gene transfer is applicable to gene medicines, such as antisense ODNs, siRNA, and adenovirus vectors. These results suggest that an Atelocollagen-based cell transfection array may be appropriate for general use in the high-throughput screening of large sets of gene medicines with functions in mammalian cells.
Honma
Kimi
K
National Cancer Center Research Institute, 1-1, Tsukiji, 5-chome, Chuo-ku, Tokyo 104-0045, Japan.
Miyata
Teruo
T
Ochiya
Takahiro
T
D003094:Collagen / Q000235:genetics*; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D005796:Genes / Q000187:drug effects*; D005822:Genetic Vectors; D006801:Humans; D010600:Pharmacology / Q000379:methods*; D014162:Transfection
C059821:atelocollagen; D003094:Collagen