Chloroquine gene transfection

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    Chloroquine gene transfection


    Detailed information on how Wiley uses cookies can be found in our Privacy Policy. This protocol describes a general method for transfecting mammalian cells using linear polyethylenimine.

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    Hours after transfection reaction has been applied to cells, gently remove media and replace with 10% glycerol or DMSO for 2-3 minutes. Remove and feed with complete media various references. *1 hour before transfection, replace cDMEM with cDMEM + 25 mM chloroquine. In gene transfection 21. TheamountofCQinpolyplexesinthe delivery of anti-miR-210 into the cancer cells, was associated with the efficient miRNA delivery as higher content of chloroquine caused increased endosomal escape 22. In another study con-ducted by Yang Hu et al. in the presence of 75 μM concentration Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred "transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant animal cells, transfection is the preferred term as.

    We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. Transfections allow for transient expression of a gene of interest in a target cell line and can be useful for short term studies of protein function.

    Chloroquine gene transfection

    Transient Transfection of 293T cells, Chloroquine a brand-new scenario for an old drug

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  7. Calcium phosphate–mediated transfection of eukaryotic cells. The uptake of DNA is markedly enhanced when the nucleic acid is presented as a coprecipitate of calcium phosphate and DNA. The insoluble precipitate attaches to the cell surface and is brought into the cells by endocytosis.

    • Calcium phosphate–mediated transfection of eukaryotic..
    • Transfection - Wikipedia.
    • Chloroquine and amphipathic peptide helices. - Gene Therapy.

    Chloroquine has been frequently used in cell culture gene transfection studies Cotton et al. 1992, Wagner et al. 1992, Lucas et al. 1995, but has not been reported yet to be successful for enabling gene transfection in vivo. As mentioned above, the mechanisms of action of chloroquine seems to be related to elevating the intravesicular pH of lysosomes and endosomes, thereby providing a suboptimal pH environment for the various lysosomal enzymes including those involved in DNA degradation. Treatments may modify cell membrane structure to enhance uptake of DNA. Chloroquine, which is generally applied to cells with the DNA, appears to enhance transfection efficiency by inhibiting DNA degradation by lysosomes 15. Sodium butyrate treatment following transfection has been reported to result in higher levels of transient expression 16. Determine DNA concentration by performing A260/A280 readings on a spectrophotometer or by using the Quant-iT™ DNA Assays Kits Q33130, Q33120. Plasmid DNA or siRNA used in transfection has degraded or is of poor quality Ensure that the plasmid DNA or siRNA used for transfection is of high quality.

     
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