Space and time-resolved gene expression experiments on cultured mammalian cells by a single-cell electroporation microarray

Single-cell experiments represent the next frontier for biochemical and gene expression research. Although bulk-scale methods averaging populations of cells have been traditionally used to investigate cellular behavior, they mask individual cell features and can lead to misleading or insufficient bi...

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Bibliographic Details
Published in:New biotechnology 2008-06, Vol.25 (1), p.55-67
Main Authors: Vassanelli, S., Bandiera, L., Borgo, M., Cellere, G., Santoni, L., Bersani, C., Salamon, M., Zaccolo, M., Lorenzelli, L., Girardi, S., Maschietto, M., Dal Maschio, M., Paccagnella, A.
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
CHO Cells
Cricetinae
Cricetulus
Electroporation - methods
Gene Expression Regulation
Gene Silencing
Green Fluorescent Proteins - metabolism
Intracellular Space - metabolism
Microelectrodes
Nucleic Acids - metabolism
Oligonucleotide Array Sequence Analysis - methods
Oligonucleotides - metabolism
RNA, Small Interfering - metabolism
Time Factors
Transfection
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Summary:Single-cell experiments represent the next frontier for biochemical and gene expression research. Although bulk-scale methods averaging populations of cells have been traditionally used to investigate cellular behavior, they mask individual cell features and can lead to misleading or insufficient biological results. We report on a single-cell electroporation microarray enabling the transfection of pre-selected individual cells at different sites within the same culture (space-resolved), at arbitrarily chosen time points and even sequentially to the same cells (time-resolved). Delivery of impermeant molecules by single-cell electroporation was first proven to be finely tunable by acting on the electroporation protocol and then optimized for transfection of nucleic acids into Chinese Hamster Ovary (CHO-K1) cells. We focused on DNA oligonucleotides (ODNs), short interfering RNAs (siRNAs), and DNA plasmid vectors, thus providing a versatile and easy-to-use platform for time-resolved gene expression experiments in single mammalian cells.
ISSN:1871-6784
1876-4347
DOI:10.1016/j.nbt.2008.03.002