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Production System was compared with polyethylenimine (PEI)–mediated transfection of lentiviral vectors in adherent HEK 293T/FT cells and suspension HEK 293 cells. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells. The resulting ...
Long-term culture and colony formation of several LV-labeled mouse melanoma cells showed that promoters derived from mammalian house-keeping genes, especially those encoding RNA polymerase II (Pol2) and ferritin (FerH), provided the highest consistency for reporter expression. For in vivo studies, ...
Both the CLIP and IDLV donors with Cas9 RNP created high GFP expression that was stable over time, indicating a targetedACTBknock-in. Inclusion of cut sites in the CLIP donor dramatically increase the percentage of K562 cells that integrate GFP into theACTBlocus, doubling the knock-in efficienc...
cells and the viral vector [105,106]. In this regard, poly-l-lysine, diethylaminoethyl (DEAE)-dextran,polybrene,protamine sulfate, or cationicliposomesare applied as positively-charged polycationic agents to diminish those repulsion forces [105,107–114]. Though utilizing polybrene inviral ...
Finally, the recombinant lentiviral vector containing miR-124 (miR-124-LV) and the recombinant lentiviral vector containing the green fluorescent protein (GFP) reporter gene (NC-EGFP-LV) were obtained. The virus titer of the cells was determined using a gradient dilution method in a 24-well ...
A image showing exceptional transfection efficiency of PolyJet™ reagent onHuman embryonic stem cells(hESCs).The hESCs grown in E8 medium on Geltrexvs(left panel, DIC imaging) was transfected with pEF1α-GFP. The GFP expression (right panel) was visualized by Nikon Eclipse Fluorescence microscope...
For example, we have studied gene-expression noise in HIV and its consequences, by utilizing lentiviral vectors expressing fluorescent reporter genes (e.g., green fluorescent protein, GFP) from the HIV-1 LTR promoter. These lentiviral vectors allowed us transduce Jurkat T cells (ATCC # TIB-152...
(MOI) of 5. GFP expression in the infected cells was observed daily under an inverted fluorescence microscope. Five days after infection, ERα recombinant lentivirus-infected and -uninfected neuronal cells were collected, and total RNA and protein were extracted. ERα expression was examined by ...
expression,FXNexpression in the human and mouse FRDA cells reached 96- and 210-fold, respectively, after infection, compared with uninfected FRDA cells 0.5- and 0.2-fold, respectively, compared with normal fibroblasts. Over time, this expression appeared to fall by ~50%, butFXNexpression ...