Transfection agents "from traditional to minaturised screening"
Transfection agents "from traditional to minaturised screening"
Three libraries of cationic lipids were synthesised and their transfection efficiency in vitro on the HEK293T cell line compared to available commercial compounds. Two compounds were shown to be more effective and showed minimal cytotoxicity by both MTT and trypan blue assays showing the value of this novel class of compounds for gene delivery in vitro.
Single bead screening was evaluated. A small library of Arginine containing cationic lipids was synthesised on high-loading beads to assess the possibility of screening the material cleaved from single beads. One compound was more efficient than EffecteneTM and seven was demonstrated transfection up to 80% efficiency. These studies showed that single bead screening was viable but further studies with accurate quantitation need to be undertaken to realise maximum activity.
Finally, several compounds from the synthesised libraries were tested for their transfection activity using a microarray setup. Poor transfection was observed, however this technique is very rapid, miniaturised and fully automated, and with further research and optimisation will offer an ideal screening technique.
University of Southampton
Yingyongnarongkul, Boon-ek
9fd5bdf6-d2f0-4b32-b1f2-72f1931d9605
2003
Yingyongnarongkul, Boon-ek
9fd5bdf6-d2f0-4b32-b1f2-72f1931d9605
Yingyongnarongkul, Boon-ek
(2003)
Transfection agents "from traditional to minaturised screening".
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Three libraries of cationic lipids were synthesised and their transfection efficiency in vitro on the HEK293T cell line compared to available commercial compounds. Two compounds were shown to be more effective and showed minimal cytotoxicity by both MTT and trypan blue assays showing the value of this novel class of compounds for gene delivery in vitro.
Single bead screening was evaluated. A small library of Arginine containing cationic lipids was synthesised on high-loading beads to assess the possibility of screening the material cleaved from single beads. One compound was more efficient than EffecteneTM and seven was demonstrated transfection up to 80% efficiency. These studies showed that single bead screening was viable but further studies with accurate quantitation need to be undertaken to realise maximum activity.
Finally, several compounds from the synthesised libraries were tested for their transfection activity using a microarray setup. Poor transfection was observed, however this technique is very rapid, miniaturised and fully automated, and with further research and optimisation will offer an ideal screening technique.
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Published date: 2003
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Local EPrints ID: 465111
URI: http://eprints.soton.ac.uk/id/eprint/465111
PURE UUID: 4e3e60ec-04e8-4580-bcce-80ac449e6f60
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Date deposited: 05 Jul 2022 00:24
Last modified: 16 Mar 2024 19:57
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Author:
Boon-ek Yingyongnarongkul
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