Transfection of cells by immunoporation

Cell transfection is now a central technique in molecular biology and an essential prerequisite for gene therapy. Here we describe how beads coated with antibodies and bound to specific cell-surface transmembrane proteins can create holes in cells when the beads are removed, allowing transfection of...

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Bibliographic Details
Published in:Nature (London) 2000-05, Vol.405 (6784), p.298-298
Main Authors: BILDIRICI, L, SMITH, P, TZAVELAS, C, HOREFTI, E, RICKWOOD, D
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
CD11b antigen
CD71 antigen
Cytological Techniques
Diverse techniques
Fundamental and applied biological sciences. Psychology
Gene transfer
Genetic engineering
Genetic technics
HL-60 Cells
Humans
Immunologic Techniques
immunoporation
Macrophage-1 Antigen - immunology
Methods. Procedures. Technologies
Microspheres
Molecular and cellular biology
Synthetic digonucleotides and genes. Sequencing
Transfection - methods
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Summary:Cell transfection is now a central technique in molecular biology and an essential prerequisite for gene therapy. Here we describe how beads coated with antibodies and bound to specific cell-surface transmembrane proteins can create holes in cells when the beads are removed, allowing transfection of the cells with DNA or other macromolecules. This unique targeted transfection of cells by immunoporation is very efficient and results in minimal cell death. A variety of methods have been developed for the transfection of cells, including electroporation, lipofection, calcium phosphate coprecipitation and DEAE dextran. Of these methods, only electroporation offers the possibility of introducing DNA and other molecules such as proteins into viable cells. None of the current methods is able to target specific types of cells for transfection. In this new method of cell transfection, antibody-coated beads are bound to specific surface antigens and then the beads are sheared off from the cell by mixing: this causes the formation of transient holes in the cell membrane through which macromolecules can enter. Granulocytic, differentiating human lymphoblastic HL-60 cells normally express CD71 on their surfaces. When induced to differentiate in the presence of dimethyl sulphoxide (DMSO), the cells cease to express CD71 and instead express CD11b. We have used this cell line as a model system to investigate the process of cell transfection mediated by immunoporation.
ISSN:0028-0836
1476-4687
DOI:10.1038/35012701