High-throughput transfection of interfering RNA into a 3D cell-culture chip
Abstract
A method for high-throughput retroviral transfection of genes and interfering RNA into 3D cell-culture microarrays is described. 3D cultures more closely mimic the in vivo cellular milieu, thus providing cellular responses to genetic manipulation more similar to the in vivo situation than 2D cultures. This technique is applied to transfect several "toxic" short-hairpin RNAs (shRNAs) into 3D cell cultures. It is demonstrated that the toxicity is similar to that obtained by conventional (non-high-throughput) retroviral transfection of cells grown in similar 3D culture microarrays.
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Organoids are embedded in extracellular matrix, which constitutes a barrier for efficient transfection.
“One limitation is that organoids are embedded in extracellular matrix and grow into compact structures that hinder transfection using traditional techniques.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [9]
Organoids grow into dense and compact structures that impede diffusion, penetration and cellular accumulation of genetic material, making transfection via traditional techniques difficult.
“Moreover, organoids grow into dense and compact structures that impede diffusion, penetration and cellular accumulation of genetic material, which makes transfection via traditional techniques difficult (7–9).”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [9]
Direct 3D transfection on already formed organoids remains challenging.
“However, direct 3D transfection on already formed organoids remains challenging.”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [9]
Lipofection and electroporation have been proven relatively inefficient in transfecting 3D cultures, with transfection efficiencies lower than 5% and 20%, respectively.
“However, they have been proven to be relatively inefficient in transfecting 3D cultures with transfection efficiencies lower than 5 and 20%, respectively (7–9).”
From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [9]
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Laperrousaz, B., Porte, S., Gerbaud, S., Härmä, V., Kermarrec, F., Hourtane, V., Bottausci, F., Gidrol, X., & Picollet-D’hahan, N. (2018) · Nucleic Acids Research · doi:10.1093/nar/gky030 · Cited by 54 · see citing works
- Authors
- Zhang, H., Lee, M., Hogg, M. G., Dordick, J. S., & Sharfstein, S. T.
- Affiliations
- Rensselaer Polytechnic Institute (US)
- Solidus Biosciences (United States) (US)
- University at Albany, State University of New York (US)
- Corresponding author
- Sharfstein, Susan T.
- Venue
- Small
- Publisher
- Wiley
- Published
- 2012-04-17
- Volume
- 8
- Issue
- 13
- Pages
- 2091-2098
- ISSN
- 1613-6810 (ISSN-L)
- 1613-6829
- OpenAlex
- W2045357269
- Cited by
- 16
- References
- 0
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2021: 1 · 2020: 2 · 2019: 1 · 2018: 2 · 2016: 2 · 2014: 1 · 2013: 2 · 2012: 2
- FWCI
- 0.64
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Small (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- retroviral transfection, interfering RNA, shRNA, 3D cell culture, gene silencing
- MeSH terms
- Animals, Cell Line, Transfection, Cell Culture Techniques, RNA Interference, NIH 3T3 Cells, Mice
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