Atelocollagen-based gene transfer in cells allows high-throughput screening of gene functions.
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.
Honma
K
K
Koken Bioscience Institute, 2-13-10, Ukima, Kita-ku, Tokyo, 115-0051, Japan.
Ochiya
T
T
Nagahara
S
S
Sano
A
A
Yamamoto
H
H
Hirai
K
K
Aso
Y
Y
Terada
M
M
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003094:Collagen; D004337:Drug Carriers; D020869:Gene Expression Profiling; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D014162:Transfection
D004337:Drug Carriers; D008164:Luminescent Proteins; D020319:Oligodeoxyribonucleotides, Antisense; C059821:atelocollagen; D049452:Green Fluorescent Proteins; D003094:Collagen