Efficiency of gene transfection reagents in NG108-15, SH-SY5Y and
CHO-K1 cell lines

Martín-Montañez, Elisa; López‐Téllez, Juan F.; Acevedo, María José; Pavía, J.; Khan, Zafar U.

doi:10.1358/mf.2010.32.5.1498327

Cited by 15 publications

(17 citation statements)

References 19 publications

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“…Lipofectamine™-2000 served as a positive control for this experiment, since use of this lipid-based transfection agent is common. In the present study, a transfection efficiency of 10%–20% was observed for Lipofectamine™-2000 (see Figure 2 ), which is comparable to the findings of other researchers who reported undifferentiated SH-SY5Y cell transfection efficiencies ranging from 0.3%–4% [ 27 , 28 ]. Furthermore, the superior performance of the nanoparticle vectors, in particular Neuromag, is revealed when compared to the established Lipofectamine™-2000 control (see Figure 2 ).…”

Section: Resultssupporting

confidence: 91%

DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons

Vernon

Dean

Dobson

2015

IJMS

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Efficient non-viral plasmid DNA transfection of most stem cells, progenitor cells and primary cell lines currently presents an obstacle for many applications within gene therapy research. From a standpoint of efficiency and cell viability, magnetic nanoparticle-based DNA transfection is a promising gene vectoring technique because it has demonstrated rapid and improved transfection outcomes when compared to alternative non-viral methods. Recently, our research group introduced oscillating magnet arrays that resulted in further improvements to this novel plasmid DNA (pDNA) vectoring technology. Continued improvements to nanomagnetic transfection techniques have focused primarily on magnetic nanoparticle (MNP) functionalization and transfection parameter optimization: cell confluence, growth media, serum starvation, magnet oscillation parameters, etc. Noting that none of these parameters can assist in the nuclear translocation of delivered pDNA following MNP-pDNA complex dissociation in the cell’s cytoplasm, inclusion of a cassette feature for pDNA nuclear translocation is theoretically justified. In this study incorporation of a DNA targeting sequence (DTS) feature in the transfecting plasmid improved transfection efficiency in model neurons, presumably from increased nuclear translocation. This observation became most apparent when comparing the response of the dividing SH-SY5Y precursor cell to the non-dividing and differentiated SH-SY5Y neuroblastoma cells.

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Section: Resultssupporting

confidence: 91%

DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons

Vernon

Dean

Dobson

2015

IJMS

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“…These fi ndings are consistent with other studies on cytotoxicity effect of Turbofect (8). Although there are many reports about the cellular toxicity of Lipofectamine 2000 in comparison with other transection reagents such as Turbofect, there are just few published studies on toxicity of Lipofectamine 3000 (9,19,(20)(21)(22)(23). In one study, Lipofectamine CRISPRMAX was compared with Lipofectamine 3000 and Lipofectamine RNAiMAX, and signifi cant effi ciency with less toxicity of Lipofectamine CRISPRMAX was shown (12,(23)(24)(25)(26).…”

Section: Discussionsupporting

confidence: 80%

Comparison of transfection efficiency of polymer-based and lipid-based transfection reagents

Rahimi¹,

Mobarakeh²,

Kamalzare³

et al. 2018

BLL

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OBJECTIVES: In this study, the optimal dose of Lipofectamine 3000 and Turbofect to transfect adherent cell lines such as CHO-K1 and HEK293 cells in comparison with non-adherent H9T-cells with pEGFP-N1 and pCDH was identifi ed. BACKGROUND: Lipofectamine 3000 is a new transfection reagent which is claimed to be more effi cient than other transfection reagents like Turbofect. Transfection effi ciency could be affected by the nature of target cell line and vector. METHODS: Transfection effi ciency and cytotoxicity of each reagent was identifi ed by using fl ow cytometry and XTT assay, respectively. RESULTS: Lipofectamine 3000 was more effi cient in transfecting pCDH, while Turbofect was more effi cient in separate transfection of CHO-K1 and HEK293 with pEGFP-N1. Lipofectamine 3000 could be cytotoxic in transfecting H9T-cells with pCDH. Also, H9T-cells were not suffi ciently transfected with each plasmid vector by using each Lipofectamine 3000 and Turbofect. Turbofect had less cytotoxicity effect on all three cell lines than Lipofectamine 3000.Transfection of suspended cells like H9T-cells by using Lipofectamine 3000 and Turbofect would not result in suffi cient transfection. CONCLUSION: Lipofectamine 3000 is the best choice for transfection of CHO-K1 and HEK293 with pCDH while Turbofect is preferably used in transfecting these cell lines with pEGFP-N1 (Tab. 1, Fig. 2, Ref. 26).

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“…The e ciency of DNA electroporation varied from about 60% for CHO-K1 to 20% for SH-SY5Y. These outcomes are comparable with previous results using lipidic transfecting agents for the delivery of a non-viral expression vector, where the high e ciency of CHO-K1 transfection (60-80%) was drastically reduced to 5% for SH-SY5Y cells [44]. Our experiments, besides demonstrating an improved e ciency for SH-SY5Y cells, paved the way for the use of in-situ capacitive electroporation for serial gene transfection of cells in adhesion.…”

Section: Discussionsupporting

confidence: 84%

Spatio-temporally targeted in-situ electroporation of mammalian cells through SiO2 thin-film capacitive microelectrodes

Maschietto

Maschio

Girardi

et al. 2020

Preprint

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Electroporation is a widely used non-viral technique for the delivery of molecules, including nucleic acids, into cells. Recently, electronic microsystems that miniaturize the electroporation machinery have been developed as a new tool for genetic manipulation of cells in vitro, by integrating metal microelectrodes in the culture substrate and enabling electroporation in-situ. We report that non-faradic SiO2 thin film-insulated microelectrodes can be used for reliable and spatially selective in-situ electroporation of mammalian cells. CHO-K1 and SH-SY5Y cell lines and primary neuronal cultures were electroporated by application of short and low amplitude voltage transients leading to cell electroporation by capacitive currents. We demonstrate reliable delivery of DNA plasmids and exogenous gene expression, accompanied by high spatial selectivity and cell viability. Finally, we show that SiO2 thin film-insulated microelectrodes support a double and serial transfection of the targeted cells.

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Efficiency of gene transfection reagents in NG108-15, SH-SY5Y and CHO-K1 cell lines

Cited by 15 publications

References 19 publications

DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons

DNA Targeting Sequence Improves Magnetic Nanoparticle-Based Plasmid DNA Transfection Efficiency in Model Neurons

Comparison of transfection efficiency of polymer-based and lipid-based transfection reagents

Spatio-temporally targeted in-situ electroporation of mammalian cells through SiO2 thin-film capacitive microelectrodes

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