Chloroquine transfection mechanism

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  1. fishika User

    Chloroquine transfection mechanism


    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.

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    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. Recently, an altered chloroquine-transporter protein CG2 of the parasite has been related to chloroquine resistance, but other mechanisms of resistance also appear to be involved. Research on the mechanism of chloroquine and how the parasite has acquired chloroquine resistance is still ongoing, as other mechanisms of resistance are likely. 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.

    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

    Transient Transfection of 293T cells, Chloroquine - Wikipedia

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    ELSEVIER BBA Biochimica et Biophysica Aqeta Biochimica et Biophysica Acta 1189 1994 195-203 DNA transfection mediated by cationic liposomes containing lipopolylysine characterization and mechanism of action Xiaohuai Zhou 1, Leaf Huang Department of Biochemistry, Graduate Program in Cell, Molecular and Developmental Biology, University of Tennessee, Knoxville, TN 37996-0840, USA Received 11. 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 efficiency was affected by the concentration of lysosomotropic agents. The marked transfection enhancement was observed when the amount of sucrose increased from 5 to 500 mM and the optimum transfection efficiency was found at 500 mM in all cell lines tested Fig. 2 a–c. Transgene expression was found to be very low when the cells were transfected with plasmid alone.

     
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