Cell Transfection

Minh, Aline Do; Sharon, David; Chahal, Parminder S.; Kamen, Amine

doi:10.1016/b978-0-444-64046-8.00023-9

Cited by 5 publications

(3 citation statements)

References 26 publications

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“…In addition, magnetic force helps penetrate the cell membrane [ 160 ]. The presence of nanoparticles near the tissue also increases the absorption of the genetic material through endocytosis [ 161 ]. It was found that such use of the magnetic field can significantly increase the efficiency of transfection of foreign genetic material into the cell [ 162 ].…”

Section: Use Of Magnetite Nanoparticles In Cancer Therapiesmentioning

confidence: 99%

Magnetite Nanoparticles in Magnetic Hyperthermia and Cancer Therapies: Challenges and Perspectives

et al. 2022

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Until now, strategies used to treat cancer are imperfect, and this generates the need to search for better and safer solutions. The biggest issue is the lack of selective interaction with neoplastic cells, which is associated with occurrence of side effects and significantly reduces the effectiveness of therapies. The use of nanoparticles in cancer can counteract these problems. One of the most promising nanoparticles is magnetite. Implementation of this nanoparticle can improve various treatment methods such as hyperthermia, targeted drug delivery, cancer genotherapy, and protein therapy. In the first case, its feature makes magnetite useful in magnetic hyperthermia. Interaction of magnetite with the altered magnetic field generates heat. This process results in raised temperature only in a desired part of a patient body. In other therapies, magnetite-based nanoparticles could serve as a carrier for various types of therapeutic load. The magnetic field would direct the drug-related magnetite nanoparticles to the pathological site. Therefore, this material can be used in protein and gene therapy or drug delivery. Since the magnetite nanoparticle can be used in various types of cancer treatment, they are extensively studied. Herein, we summarize the latest finding on the applicability of the magnetite nanoparticles, also addressing the most critical problems faced by smart nanomedicine in oncological therapies.

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Section: Use Of Magnetite Nanoparticles In Cancer Therapiesmentioning

confidence: 99%

Magnetite Nanoparticles in Magnetic Hyperthermia and Cancer Therapies: Challenges and Perspectives

et al. 2022

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“…TRs are designed to facilitate the introduction of nucleic acids into eukaryotic cells and usually feature a positively charged head group and one or two hydrocarbon chains that bind to negatively charged nucleic acids via electrostatic interactions to form cationic complexes (also known as lipoplexes). These complexes interact with the negatively charged phospholipid bilayer of the cell membrane, promoting uptake and intracellular release [ 36 – 39 ]. The formulation and composition of most commercially available TRs is not disclosed [ 40 ], making it difficult to rationally select TRs based on their components.…”

Section: Introductionmentioning

confidence: 99%

Gene silencing in the aedine cell lines C6/36 and U4.4 using long double-stranded RNA

Omokungbe,

Centurión,

Stiehler

et al. 2024

Parasites Vectors

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Background RNA interference (RNAi) is a target-specific gene silencing method that can be used to determine gene functions and investigate host–pathogen interactions, as well as facilitating the development of ecofriendly pesticides. Commercially available transfection reagents (TRs) can improve the efficacy of RNAi. However, we currently lack a product and protocol for the transfection of insect cell lines with long double-stranded RNA (dsRNA). Methods We used agarose gel electrophoresis to determine the capacity of eight TRs to form complexes with long dsRNA. A CellTiter-Glo assay was then used to assess the cytotoxicity of the resulting lipoplexes. We also measured the cellular uptake of dsRNA by fluorescence microscopy using the fluorophore Cy3 as a label. Finally, we analyzed the TRs based on their transfection efficacy and compared the RNAi responses of Aedes albopictus C6/36 and U4.4 cells by knocking down an mCherry reporter Semliki Forest virus in both cell lines. Results The TRs from Biontex (K4, Metafectene Pro, and Metafectene SI+) showed the best complexing capacity and the lowest dsRNA:TR ratio needed for complete complex formation. Only HiPerFect was unable to complex the dsRNA completely, even at a ratio of 1:9. Most of the complexes containing mCherry-dsRNA were nontoxic at 2 ng/µL, but Lipofectamine 2000 was toxic at 1 ng/µL in U4.4 cells and at 2 ng/µL in C6/36 cells. The transfection of U4.4 cells with mCherry-dsRNA/TR complexes achieved significant knockdown of the virus reporter. Comparison of the RNAi response in C6/36 and U4.4 cells suggested that C6/36 cells lack the antiviral RNAi response because there was no significant knockdown of the virus reporter in any of the treatments. Conclusions C6/36 cells have an impaired RNAi response as previously reported. This investigation provides valuable information for future RNAi experiments by showing how to mitigate the adverse effects attributed to TRs. This will facilitate the judicious selection of TRs and transfection conditions conducive to RNAi research in mosquitoes. Graphical Abstract

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“…Magnetic beads are also used for transfection in a process known as magnetofection. Iron oxide particles complexed with nucleic acids are forced into the cells with a strong magnetic field resulting in close contact and subsequent endocytosis [43,44].…”

Section: Introductionmentioning

confidence: 99%

Lipofection mediated transfection fails for sea urchin coelomocytes

Hudgell

Smith

2022

PLoS ONE

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Molecular cloning, gene manipulation, gene expression, protein function, and gene regulation all depend on the introduction of nucleic acids into target cells. Multiple methods have been developed to facilitate such delivery including instrument based microinjection and electroporation, biological methods such as transduction, and chemical methods such as calcium phosphate precipitation, cationic polymers, and lipid based transfection, also known as lipofection. Here we report attempts to lipofect sea urchin coelomocytes using DOTAP lipofection reagent packaged with a range of molecules including fluorochromes, in addition to expression constructs, amplicons, and RNA encoding GFP. DOTAP has low cytotoxicity for coelomocytes, however, lipofection of a variety of molecules fails to produce any signature of success based on results from fluorescence microscopy and flow cytometry. While these results are negative, it is important to report failed attempts so that others conducting similar research do not repeat these approaches. Failure may be the outcome of elevated ionic strength of the coelomocyte culture medium, uptake and degradation of lipoplexes in the endosomal-lysosomal system, failure of the nucleic acids to escape the endosomal vesicles and enter the cytoplasm, and difficulties in lipofecting primary cultures of phagocytic cells. We encourage others to build on this report by using our information to optimize lipofection with a range of other approaches to work towards establishing a successful method of transfecting adult cells from marine invertebrates.

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Cell Transfection

Cited by 5 publications

References 26 publications

Magnetite Nanoparticles in Magnetic Hyperthermia and Cancer Therapies: Challenges and Perspectives

Magnetite Nanoparticles in Magnetic Hyperthermia and Cancer Therapies: Challenges and Perspectives

Gene silencing in the aedine cell lines C6/36 and U4.4 using long double-stranded RNA

Lipofection mediated transfection fails for sea urchin coelomocytes

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