304. Atelocollagen-Based Cell Transfection Array Allows High-Throughput Screening of Gene Functions
The human genome project has been completed, and it is urgently required that genome information has been implicated in drug discovery, gene therapy and regenerative medicine. To achieve this goal, we have developed cell-based gene expression array that can elucidate the function of a large number o...
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| Published in: | Molecular therapy Vol. 9; no. S1; pp. S116 - S117 |
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| Main Authors: | , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Milwaukee
Elsevier Inc
01-05-2004
Elsevier Limited |
| Subjects: | |
| Online Access: | Get full text |
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| Summary: | The human genome project has been completed, and it is urgently required that genome information has been implicated in drug discovery, gene therapy and regenerative medicine. To achieve this goal, we have developed cell-based gene expression array that can elucidate the function of a large number of genes within a short time, designated Atelocollagen-based cell transfection array. It is essential to employ materials 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, biocompatible and low immunogenic biomaterial, is a promising carrier for gene delivery because it is considered safe and appropriate for practical use. We have successfully used Atelocollagen for high-throughput gene transfer into cells and expression screening of gene function. In this system, Atelocollagen/genetic material complexes with a nano-size particle are pre-coated on a micro-well plate on which the cells are then seeded. Through this method, the cells take up the genetic materials and express them by reverse transfection. Atelocollagen/genetic complexes allow for long-term gene expression and are free from cytotoxicity. Then, the cellular biology of the transduced cells could be studied at once with multiparameters including cell growth, cell morphology, subcellular localization of the protein products, number of apoptotic cells, and up- and down-regulation of signaling molecules by using conventional assay systems. We confirmed that the present method with Atelocollagen-based gene transfer is applicable to gene medicines, such as plasmid DNAs, adenovirus vectors, antisense oligonucleotides and siRNAs, and to various cells for cell-based assays.Here, we present an experimental demonstration of an Atelocollagen-based cell transfection array for the detection of genes with neurite-extension ability from large sets of cDNA libraries of developmental mice brain in PC12 cells, rat adrenal pheochromocytoma. Furthermore, siRNAs which have become a powerful tool for gene knockdown were utilized to characterize the genes participating in drug resistance in mammalian cells.These results suggest that an Atelocollagen-based cell transfection array may allow high-throughput screening of large sets of gene medicines with functions in mammalian cells and should greatly contribute to drug discovery, gene therapy and regenerative medicine. |
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| ISSN: | 1525-0016 1525-0024 |
| DOI: | 10.1016/j.ymthe.2004.06.246 |