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. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. Puckering of tongue plaquenil Hydroxychloroquine is 200 mg enough to take Chloroquine and release of cargo Chloroquine is a member of quinolone family and is a weak intercalating agent. Chloroquine is used for treating amebiasis, rheumatoid arthritis, discoid and systemic lupus erythematosus. Application DNA intercalator. Also used to increase transfection efficiency. Chloroquine diphosphate salt has been used • in in vitro antiplasmodial assays Chloroquine has been used to increase transfection efficiency for calcium phosphate, DEAE-dextran, and some liposome-mediated gene delivery 63,65–68. However, effects of lysosomotropic agents on polymer and some lipid based methods of gene delivery are still controversial in the literature. Similarly, the transformation frequency of rat cells following polyoma DNA transfection was approximately 6-fold increased by chloroquine treatment. The effect of the compound was even more pronounced in transfections with linear forms of polyoma DNA, suggesting that chloroquine inhibits degradation of DNA absorbed by the cells. Last Upload: June 10, 2016 Day 0: Seed Lenti-X 293T cells (this cell line is optimized for production of lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment *Pro-Tips* Different brands and lots of FBS can promote or inhibit transfection. This approach can be adapted for different cell lines and different transfection reagents. Chloroquine transfection mechanism Dictyostelium Discoideum Expresses a Malaria Chloroquine., Distinct effects of endosomal escape and inhibition of endosomal. Plaquenil special considerations Figure 1. Structure of chloroquine and some related synthetic compounds that are effective against chloroquine-resistant as well as chloroquine-sensitive P. binding depends on the stacking interactions between the 7-chloro-substituted quinoline ring and products of heme released from the digestion of host cell hemoglobin. Plasmodium Chloroquine Resistance and the Search for a Replacement.. High efficiency polyoma DNA transfection of chloroquine treated cells.. DNA transfection mediated by cationic liposomes containing.. Carefully transfer the transfection mix to the Lenti-X 293T packaging cells. Add the transfection mix dropwise being careful not to dislodge the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully aspirate the media. Replace the media with 15 mL of DMEM complete. Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer technology enables the study of gene function and protein expression in a cellular environment. To understand how chloroquine CQ enhances transgene expression in polycation-based, nonviral gene delivery systems, a number of CQ analogues with variations in the aliphatic amino side chain or in the aromatic ring are synthesized and investigated.